This invention relates to sodium-hydrogen exchanger type 1 (NHE-1) inhibitors, pharmaceutical compositions containing such inhibitors and the use of such inhibitors to treat for example, ischemia particularly, perioperative myocardial ischemic injury in mammals, including humans.
Mycardial ischemic injury can occur in out-patient as well as in perioperative settings and can lead to the development of sudden death, myocardial infarction or congestive heart failure. There is an unmet medical need to prevent or minimize myocardial ischemic injury, particularly perioperative myocardial infarction. Such a therapy is anticipated to be life-saving and reduce hospitalizations, enhance quality of life and reduce overall health care costs of high risk patients.
Pharmacological cardioprotection would reduce the incidence and progression of myocardial infarction and dysfunction occurring in these surgical settings (perioperatively). In addition to reducing myocardial damage and improving post-ischemic myocardial function in patients with ischemic heart disease, cardioprotection would also decrease the incidence of cardiac morbidity and mortality due to myocardial infarction and dysfunction in patients xe2x80x9cat riskxe2x80x9d (such as greater than 65 years, exercise intolerant, coronary artery disease, diabetes mellitus, hypertension) that require non-cardiac surgery.
The mechanism(s) responsible for the myocardial injury observed after ischemia and reperfusion is not fully understood.
A variety of publications have disclosed the use of guanidine derivatives as useful for the treatment of, for example arrhythmias.
U.S. Pat. No. 5,698,581, granted Dec. 16, 1997 (EP 676395 A2 published 1995), discloses certain substituted N-heteroarylguanidines as inhibitors of the (Na+/H+) exchange transport system useful for the treatment of, for example, arrhythmias.
EP 803 501 A1, published Oct. 10, 1997, discloses substituted guanidine derivatives useful as (Na+/H+) exchange inhibitors.
WO 94/26709 discloses guanidine derivatives as inhibitors of (Na+/H+) exchange in cells.
PCT/JP97/04650 application published on Jun. 25, 1998 discloses N-[(substituted five-membered heteroaryl)carbonyl]guanidine compounds which are stated to be useful as inhibitors of Na+/H+ exchange and consequently effective for the treatment of various diseases such as hypertension, arrhythmia, angina pectoris, myocardial infarct, arteriosclerosis, and complications of diabetes.
Thus, there is clearly a need and a continuing search in this field of art for treatments for perioperative myocardial ischemia.
This invention is directed to a compound of Formula I 
a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug wherein
Z is carbon connected and is a five-membered, diaza, diunsaturated ring having two contiguous nitrogens, said ring optionally mono-, di-, or tri-substituted with up to three substituents independently selected from R1, R2 and R3;
or
Z is carbon connected and is a five-membered, triaza, diunsaturated ring, said ring optionally mono- or di-substituted with up to two substituents independently selected from R4 and R5;
wherein R1, R2, R3, R4 and R5 are each independently hydrogen, hydroxy(C1-C4)alkyl, (C1-C4)alkyl, (C1-C4)alkylthio, (C3-C4)cycloalkyl, (C3-C7)cycloalkyl(C1-C4)alkyl, (C1-C4)alkoxy, (C1-C4)alkoxy(C1-C4) alkyl, mono-N-xe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, M or M(C1-C4)alkyl, any of said previous (C1-C4)alkyl moieties optionally having from one to nine fluorines; said (C1-C4)alkyl or (C3-C4)cycloalkyl optionally mono- or di-substituted independently with hydroxy, (C1-C4)alkoxy, (C1-C4)alklthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, (C1-C4)alkyl, mono-N- or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl; and said (C3-C4)cycloalkyl optionally having from one to seven fluorines;
wherein M is a partially saturated, fully saturated or fully unsaturated five to eight membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen, or, a bicyclic ring consisting of two fused partially saturated, fully saturated or fully unsaturated three to six membered rings, taken independently, optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen;
said M is optionally substituted, on one ring if the moiety is monocyclic, or one or both rings if the moiety is bicyclic, on carbon or nitrogen with up to three substituents independently selected from R6, R7 and R8, wherein one of R6, R7 and R8 is optionally a partially saturated, fully saturated, or fully unsaturated three to seven membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen optionally substituted with (C1-C4)alkyl and additionally R6, R7 and R8 are optionally hydroxy, nitro, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, formyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl, (C2-C4)alkenyl, (C2-C4)alkynyl or (C5-C7)cycloalkenyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino or (C3-C7)cycloalkyl R6, R7 and R8 substituents are optionally mono-substituted independently with hydroxy, (C1-C4)alkoxycarbonyl, (C3-C7)cycloalkyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido; (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4) alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, nitro, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines.
A preferred group of compounds, designated the A group, contains those compounds having the Formula I as shown above wherein Z is 
R1 and R3 are each independently hydrogen, (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R1 and R3 substituents optionally mono- or di-subtituted independently with hydroxy, (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R2 is unsubstituted (C1-C4)alkyl or (C3-C7)cycloalkyl; or R2 is phenyl, phenyl(C1-C4)alkyl, pyridyl or pyrimidinyl or a bicyclic ring consisting of two fused five and/or six membered rings taken independently optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R2 substituent optionally mono-, di- or tri-substituted independently with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, (C1-C4)alkoxycarbonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, (C1-C4)alkylsulfonyl or sulfonamido, said (C1-C4)alkyl or (C1-C4)alkoxy optionally substituted with from one to nine fluorines or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the A Group of compounds designated the B Group, contains those compounds wherein
R1 is (C1-C4)alkyl or (C3-C7)cycloalkyl;
R2 is phenyl, optionally mono- or di-substituted; and
R3 is hydrogen or the pharmaceutically acceptable salts thereof.
Especially preferred compounds of Formula I are the compounds
[1-(2-chlorophenyl)-5-methyl-1H-pyrazole-4-carbonyl]guanidine;
[5-methyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carbonyl]guanidine;
[5-ethyl-1-phenyl-1H-pyrazole-4-carbonyl]guanidine;
[5-cyclopropyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carbonyl]guanidine;
[5-cyclopropyl-1-phenyl-1H-pyrazole-4-carbonyl]guanidine;
[5-cyclopropyl-1-(2,6-dichlorophenyl)-1H-pyrazole-4-carbonyl]guanidine and the pharmaceutically acceptable salts thereof.
Especially preferred compounds within the B Group of compounds are compounds wherein
a.
R2 is 2-chlorophenyl; and
R1 is methyl;
b.
R2 is 2-trifluoromethylphenyl; and
R1 is methyl;
c.
R2 is phenyl; and
R1 is ethyl;
d.
R2 is 2-trifluoromethylphenyl; and
R1 is cyclopropyl;
e.
R2 is phenyl; and
R1 is cyclopropyl; and
f.
R2 is 2,6-dichlorophenyl; and
R1 is cyclopropyl or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the A Group of compounds designated the C Group, contains those compounds wherein
R1 is (C1-C4)alkyl or (C3-C7)cycloalkyl;
R2 is naphthalenyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinoxalinyl, quinazolinyl, benzopyranyl, benzothiophenyl, benzodioxanyl or benzodioxolyl, said R2 substituent optionally mono-substituted; and
R3 is hydrogen or the pharmaceutically acceptable salts thereof.
Especially preferred compounds of Formula I are the compounds
[5-methyl-1-(quinolin-6-yl)-1H-pyrazole-4-carbonyl]guanidine;
[5-methyl-1-(naphthalen-1-yl)-1H-pyrazole-4-carbonyl]guanidine;
[5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carbonyl]guanidine;
[5-cyclopropyl-1-(quinolin-8-yl)-1H-pyrazole-4-carbonyl]guanidine and the pharmaceutically acceptable salts thereof.
Especially preferred compounds within the C Group of compounds are compounds wherein
a.
R2 is 1-naphthalenyl; and
R1 is methyl;
b.
R2 is 5-quinolinyl; and
R1 is cyclopropyl;
c.
R2 is 8-quinolinyl; and
R1 is cyclopropyl; and
d.
R2 is 6-quinolinyl; and
R1 is methyl or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the A Group of compounds, designated the D Group, contains those compounds wherein
R1 is hydrogen;
R2 is phenyl, optionally mono- or di-substituted; and
R3 is (C1-C4)alkyl or (C3-C7)cycloalkyl or the pharmaceutically acceptable salts thereof.
Especially preferred compounds of Formula I are the compounds
[3-methyl-1-phenyl-1H-pyrazole-4-carbonyl]guanidine;
[3-methyl-1-(naphthalen-1-yl)-1H-pyrazole-4-carbonyl]guanidine;
[3-methyl-1-(isoquinolin-5-yl)-1H-pyrazole-4-carbonyl]guanidine and the pharmaceutically acceptable salts thereof.
An especially preferred compound within the D Group of compounds is the compound wherein
R2 is phenyl; and
R3 is methyl or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the A Group of compounds, designated the E Group, contains those compounds wherein
R1 is hydrogen;
R2 is naphthalenyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinoxalinyl, quinazolinyl, benzopyranyl, benzothiophenyl, benzodioxanyl or benzodioxolyl, said R2 substituent optionally mono-substituted; and
R3 is (C1-C4)alkyl or (C3-C7)cycloalkyl or the pharmaceutically acceptable salts thereof.
Especially preferred compounds within the E Group of compounds are compounds wherein
a.
R2 is 1-naphthalenyl; and
R3 is methyl; and
b.
R2 is 5-isoquinolyl; and
R3 is methyl or the pharmaceutically acceptable salts thereof.
A preferred group of compounds, designated the F Group, contains those compounds having the Formula I as shown above wherein Z is 
R1 is hydrogen, (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R1 substituents optionally mono- or di-substituted independently with hydroxy, (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R2 and R3 are each independently unsubstituted (C1-C4)alkyl or (C3-C7)cycloalkyl; or R2 and R3 are each independently phenyl or phenyl(C1-C4)alkyl, pyridyl or pyrimidinyl or a bicyclic ring consisting of two fused five and/or six membered rings taken independently optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R2 and R3 substituents optionally mono-, di- or tri-substituted independently with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, trifluoromethoxy, (C1-C4)alkoxycarbonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, mono-Nxe2x80x94 or di-N,N-C1-C4)alkylamino, (C1-C4)alkylsulfonyl or sulfonamido, said (C1-C4)alkyl or (C1-C4)alkoxy optionally substituted with from one to nine fluorines or the pharmaceutically acceptable salts thereof.
An especially preferred compound of Formula I is
[4-methyl-1-phenyl-1H-pyrazole-3-carbonyl]guanidine and a pharmaceutically acceptable salt thereof.
An especially preferred compound within the F Group of compounds is a compound wherein
R3 is phenyl;
R1 is methyl; and
R2 is H or the pharmaceutically acceptable salts thereof.
A preferred group of compounds, designated the G Group, contains those compounds having the Formula I as shown above wherein Z is 
R1 and R3 are each independently hydrogen, (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R1 and R3 substituents optionally mono- or di-substituted independently with hydroxy, (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R2 is unsubstituted (C1-C4)alkyl or (C3-C7)cycloalkyl; or R2 is phenyl, phenyl(C1-C4)alkyl, pyridyl or pyrimidinyl or a bicyclic ring consisting of two fused five and/or six membered rings taken independently optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R2 substituent optionally mono-, di- or tri-substituted independently with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, (C1-C4)alkoxycarbonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94C1-C4)alkylamino, (C1-C4)alkylsulfonyl or sulfonamido, said (C1-C4)alkyl or (C1-C4)alkoxy optionally substituted with from one to nine fluorines or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the G Group of compounds, designated the H Group, contains those compounds wherein
R1 is (C1-C4)alkyl or (C3-C7)cycloalkyl;
R2 is phenyl, optionally mono- or di-substituted; and
R3 is hydrogen or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the G Group of compounds, designated the I Group, contains those compounds wherein
R1 is (C1-C4)alkyl or (C3-C7)cydoalkyl;
R2 is naphthalenyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinoxalinyl, quinazolinyl, benzopyranyl, benzothiophenyl, benzodioxanyl or benzodioxolyl, said R2 substituent optionally mono-substituted; and
R3 is hydrogen.
A group of compounds which is preferred among the G Group of compounds, designated the J Group, contains those compounds wherein
R1 is hydrogen;
R2 is phenyl, optionally mono- or di-substituted; and
R3 is (C1-C4)alkyl or (C3-C7)cycloalkyl.
Especially preferred compounds of Formula I are the compounds
[2-methyl-5-phenyl-2H-pyrazole-3-carbonyl]guanidine;
[2-methyl-5-(naphthalen-1-yl)-2H-pyrazole-3-carbonyl]guanidine and the pharmaceutically acceptable salts thereof.
An especially preferred compound within the J Group of compounds is the compound wherein
R2 is phenyl; and
R3 is methyl or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the G Group of compounds, designated the K Group, contains those compounds wherein
R1 is hydrogen;
R2 is naphthalenyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinoxalinyl, quinazolinyl, benzopyranyl, benzothiophenyl, benzodioxanyl or benzodioxolyl, said R2 substituent optionally mono-substituted; and
R3 is (C1-C4)alkyl or (C3-C7)cycloalkyl or the pharmaceutically acceptable salts thereof.
An especially preferred compound within the K Group of compounds is the compound wherein
R2 is 1-naphthalenyl; and
R3 is methyl and the pharmaceutically acceptable salts thereof.
A preferred group of compounds, designated the L Group, contains those, compounds having the Formula I as shown above wherein Z is 
R4 is hydrogen, (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R4 substituent optionally mono- or di-substituted independently with hydroxy, (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1C4)alkylsulfonyl; and
R5 is unsubstituted (C1-C4)alkyl or (C3-C7)cycloalkyl; or R5 is phenyl, phenyl(C1-C4)alkyl, pyridyl or pyrimidinyl or a bicyclic ring consisting of two fused five and/or six membered rings taken independently optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R5 substituent optionally mono-, di- or tri-substituted independently with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, (C1-C4)alkoxycarbonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, (C1-C4)alkylsulfonyl, or sulfonamido, said (C1-C4)alkyl or (C1-C4)alkoxy optionally substituted with from one to nine fluorines or the pharmaceutically acceptable salts thereof.
A preferred group of compounds, designated the M Group, contains those compounds having the Formula I as shown above wherein Z is 
R4 is hydrogen, (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl, phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R4 substituent optionally mono- or di-substituted independently with hydroxy, (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R5 is unsubstituted (C1-C4)alkyl or (C3-C7)cycloalkyl; or R5 is phenyl, phenyl(C1-C4)alkyl, pyridyl or pyrimidinyl or a bicyclic ring consisting of two fused five and/or six membered rings taken independently optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R5 substituent optionally mono-, di- or tri-substituted independently with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, (C1-C4)alkoxycarbonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, (C1-C4)alkylsulfonyl or sulfonamido, said (C1-C4)alkyl or (C1-C4)alkoxy optionally substituted with from one to nine fluorines or the pharmaceutically acceptable salts thereof.
A preferred group of compounds, designated the N Group, contains those compounds having the Formula I as shown above wherein Z is 
R4 is hydrogen, (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R4 substituent optionally mono- or di-substituted independently with hydroxy, (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R5 is unsubstituted (C1-C4)alkyl or (C3-C7)cycloalkyl; or R5 is phenyl, phenyl(C1-C4)alkyl, pyridyl or pyrimidinyl or a bicyclic ring consisting of two fused five and/or six membered rings taken independently optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R5 substituent optionally mono-, di- or tri-substituted independently with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, (C1-C4)alkoxycarbonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, (C1-C4)alkylsulfonyl or sulfonamido, said (C1-C4)alkyl or (C1-C4)alkoxy optionally substituted with from one to nine fluorines or the pharmaceutically acceptable salts thereof.
A preferred group of compounds, designated the O Group, contains those compounds having the Formula I as shown above wherein Z is 
R4 is hydrogen, (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R4 substituent optionally mono- or di-substituted independently with hydroxy, (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R5 is unsubstituted (C1-C4)alkyl or (C3-C7)cycloalkyl; or R5 is phenyl, phenyl(C1-C4)alkyl, pyridyl or pyrimidinyl or a bicyclic ring consisting of two fused five and/or six membered rings taken independently optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R5 substituent optionally mono-, di- or tri-substituted independently with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, (C1-C4)alkoxycarbonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, (C1-C4)alkylsulfonyl or sulfonamido, said (C1-C4)alkyl or (C1-C4)alkoxy optionally substituted with from one to nine fluorines or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the O Group of compounds designated the P Group, contains those compounds wherein
R4 is (C1-C4)alkyl or (C3-C7)cycloalkyl; and
R5 is phenyl, optionally mono- or di-substituted or the pharmaceutically acceptable salts thereof.
Especially preferred compounds of Formula I are the compounds
[5-methyl-2-phenyl-2H-1,2,3-triazole-4-carbonyl]guanidine;
[5-methyl-2-(3-methoxyphenyl)-2H-1,2,3-triazole-4-carbonyl]guanidine;
[2-(3-bromophenyl)-5-methyl-2H-1,2,3-triazole-4-carbonyl]guanidine and the pharmaceutically acceptable salts thereof.
Especially preferred compounds within the P Group of compounds are compounds wherein
a.
R5 is phenyl; and
R4 is methyl;
b.
R5 is 3-methoxyphenyl; and
R4 is methyl; and
c.
R5 is 3-bromophenyl; and
R4 is methyl or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the O Group of compounds, designated the Q Group, contains those compounds wherein
R4 is (C1-C4)alkyl or (C3-C7)cycloalkyl; and
R5 is naphthalenyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinoxalinyl, quinazolinyl, benzopyranyl, benzothiophenyl, benzodioxanyl or benzodioxolyl, said R5 substituents optionally mono-substituted or the pharmaceutically acceptable salts thereof.
Especially preferred compounds of Formula I are the compounds
[2-(naphthalen-1-yl)-5-methyl-2H-1,2,3-triazole-4-carbonyl]guanidine;
[2-(isoquinolin-5-yl)-5-methyl-2H-1,2,3-triazole-4-carbonyl]guanidine;
[5-methyl-2-(quinolin-5-yl)-2H-1,2,3-triazole-4-carbonyl]guanidine and the pharmaceutically acceptable salts thereof.
Especially preferred compounds within the Q Group of compounds are compounds wherein
a.
R5 is 1-naphthalenyl; and
R4 is methyl;
b.
R5 is 5-isoquinolinyl; and
R4 is methyl; and
c.
R5 is 5-quinolinyl; and
R4 is methyl or the pharmaceutically acceptable salts thereof.
Another aspect of this invention is directed to the following compounds:
5-Methyl-2-(5-quinolinyl)-2H-1,2,3-triazole-4-carboxylic acid,
5-Methyl-2-(5-isoquinolinyl)-2H-1,2,3-triazole-4-carboxylic acid,
2-(1-Naphthalenyl)-5-methyl-2H-1,2,3-triazole-4-carboxylic acid,
Ethyl 5-cyclopropyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carboxylate,
Ethyl 5-methyl-1-(6-quinolinyl)-1H-pyrazole-4-carboxylate,
Ethyl 5-methyl-1-naphthalenyl-1H-pyrazole-4-carboxylate,
Ethyl 5-cyclopropyl-1-(quinolin-8-yl)-1H-pyrazole-4-carboxylate,
Ethyl 5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carboxylate,
Methyl 5-ethyl-1-(quinolin-5-yl)-1H-pyrazole-4-carboxylate,
n-Butyl 1-(isoquinolin-5-yl)-3-methyl-1H-pyrazole-4-carboxylate,
5-Methyl-1-(6-quinolinyl)-1H-pyrazole-4-carboxylic acid,
5-Methyl-1-naphthalenyl-1H-pyrazole-4-carboxylic acid,
5-Cyctlopropyl-1-(quinolin-8-yl)-1H-pyrazole-4-carboxylic acid,
5-Cyclopropyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carboxylic acid,
5-Ethyl-1-(quinolin-5-yl)-1H-pyrazole-4-carboxylic acid,
5-Cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carboxylic acid or
1-(Isoquinolin-5-yl)-3-methyl-1H-pyrazole-4-carboxylic acid or a pharmaceutically acceptable salt of said compound.
A preferred group of compounds designated the R group, contains those compounds having the Formula I as shown above wherein Z is 
R1 is (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C3-C7)cycloalkyl optionally substituted with from one to three fluorines, said R1 substituent optionally mono- or di-substituted independently with (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R2 is (C1-C4)alkyl, (C3-C4)cycloalkyl, M or M(C1-C4)alkyl, any of said previous (C1-C4)alkyl moieties optionally having from one to nine fluorines; said (C1-C4)alkyl or (C3-C4)cycloalkyl optionally mono- or di-substituted independently with hydroxy, (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, (C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl; and said (C3-C4)cycloalkyl optionally having from one to seven fluorines;
wherein M is a partially saturated, fully saturated or fully unsaturated five to eight membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen, or, a bicyclic ring consisting of two fused partially saturated, fully saturated or fully unsaturated three to six membered rings, taken independently, optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen;
said M is optionally substituted, on one ring if the moiety is monocyclic, or one or both rings if the moiety is bicyclic, on carbon or nitrogen with up to three substituents independently selected from R6, R7 and R8, wherein one of R6, R7 and R8 is optionally a partially saturated, fully saturated, or fully unsaturated three to seven membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen optionally substituted with (C1-C4)alkyl and additionally R6, R7 and R8 are optionally hydroxy, nitro, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, formyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono,Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl, (C2-C4)alkenyl, (C2-C4)alkynyl or (C5-C7)cycloalkenyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alitylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino or (C3-C7)cycloalkyl R6, R7 and R8 substituents are optionally mono-substituted independently with hydroxy, (C1-C4)alkoxycarbonyl, (C3-C7)cycloalkyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfon, amido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, nitro, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines or the pharmaceutically acceptable salts thereof.
An especially preferred compound of Formula I is the compound
[1-(Naphthalen-1-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the R Group of compounds designated the S Group, contains those compounds wherein
R1 is cyclopropyl; and
R2 is 1-naphthalenyl or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the R Group of compounds designated the T Group, contains those compounds wherein
R1 is (C3-C7)cycloalkyl; and
R2 is a five to six membered monocyclic aromatic ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen;
said R2 ring is optionally mono-substituted on carbon or nitrogen with a fully saturated or fully unsaturated five to six membered ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen, said ring optionally mono-substituted with (C1-C4)alkyl
said R2 ring is also optionally mono- or di-substituted independently on carbon or nitrogen with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino are optionally mono-substituted with hydroxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfornamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the T Group of compounds designated the U Group, contains those compounds wherein
R1 is cyclopropyl; and
R2 is phenyl, optionally mono- or di-substituted independently with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino substituents are optionally mono-substituted with hydroxy, (C1-C4)alkanoylamino, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to five fluorines;
or the pharmaceutically acceptable salts thereof.
Especially preferred compounds, of Formula I are the compounds
[5-cyclopropyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carbonyl]guanidine;
[5-cyclopropyl-1-phenyl-1H-pyrazole-4-carbonyl]guanidine; or
[5-cyclopropyl-1-(2,6-dichlorophenyl)-1H-pyrazole-4-carbonyl]guanidine
or the pharmaceutically acceptable salts of said compounds.
Other especially preferred compounds of Formula I are the compounds
[1-(2-Chloro-4-methylsulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Chlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Trifluoromethyl-4-fluorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Bromophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Fluorophenyl-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Chloro-5-methoxyphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Chloro-4-methylaminosulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2,5-Dichlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2,3-Dichlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Chloro-5-aminocarbonylphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Chloro-5-aminosulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Fluoro-6-trifluoromethylphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Chloro-5-methylsulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Chloro-5-dimethylaminosulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Trifluoromethyl-4-chlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guahidine;
or pharmaceutically acceptable salts of said compounds.
Especially preferred compounds within the U Group are compounds wherein
a. R2 is 2-chloro-4-methylsulfonylphenyl;
b. R2 is 2-chlorophenyl;
c. R2 is 2-trifluoromethyl-4-fluorophenyl;
d. R2 is 2-bromophenyl;
e. R2 is 2-fluorophenyl;
f. R2 is 2-chloro-5-methoxyphenyl;
g. R2 is 2-chloro-4-methylaminosulfonylphenyl;
h. R2 is 2,5-dichlorophenyl;
i. R2 is 2,3-dichlorophenyl;
j. R2 is 2-chloro-5-aminocarbonylphenyl;
k. R2 is 2-chloro-5-aminosulfonylphenyl;
l. R2 is 2-fluoro-6-trifluoromethylphenyl;
m. R2 is 2-chloro-5-methylsulfonylphenyl;
n. R2 is 2-chloro-5-dimethylaminosulfonylphenyl;
o. R2 is 2-trifluoromethyl-4-chlorophenyl; or the pharmaceutically acceptable salts of said compounds.
A group of compounds which is preferred among the R Group of compounds designated the W Group, contains those compounds wherein
R2 is a five to six membered nonaromatic heterocyclic ring having one to two heteroatoms selected independently from nitrogen, sulfur and oxygen or R2 is unsubstituted (C1-C4)alkyl, unsubstituted (C3-C7)cycloalkyl or phenyl(C1-C4)alkyl, wherein said phenyl(C1-C4)alkyl is optionally mono- or di-substituted independently with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamirto substituents are optionally mono-substituted with hydroxy, (C1-C4)alkanoylamino, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to five fluorines;
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the R Group of compounds designated the X Group, contains those compounds wherein
R2 is a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R2 substituent optionally substituted on carbon or nitrogen with up to three substituents independently selected from R6, R7 and R8, wherein one of R6, R7 and R8 is optionally a partially saturated, fully saturated, or fully unsaturated three to seven membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen optionally substituted with (C1-C4)alkyl and additionally R6, R7 and R8 are optionally hydroxy, nitro, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, formyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl, (C2-C4)alkenyl, (C2-C4)alkynyl or (C5-C7)cycloalkenyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino or (C3-C7)cycloalkyl R6, R7 and R8 substituents are optionally mono-substituted independently with hydroxy, (C1-C4)alkoxycarbonyl, (C3-C7)cycloalkyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, nitro, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,N(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines,
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the X Group of compounds designated the Y Group, contains those compounds wherein
R1 is (C3-C7)cycloalkyl; and
R2 is a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently having one to three heteroatoms selected independently from nitrogen, sulfur and oxygen,
said R2 bicyclic ring is optionally mono-substituted on carbon or nitrogen with a fully saturated or fully unsaturated five to six membered ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen, said ring optionally mono-substituted with (C1-C4)alkyl
said R2 bicyclic ring is also optionally mono- or di-substituted independently on carbon or nitrogen with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino are optionally mono-substituted with hydroxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the Y Group of compounds designated the Z Group, contains those compounds wherein
R1 is cyclopropyl; and
R2 is a quinazolinyl, phthalazinyl, quinolinyl, isoquinolinyl, cinnolinyl, benzodioxanyl, quinoxalinyl, benzopyranyl, benzothiophenyl, benzodioxolyl, benzirmidazolyl, indazolyl, indolyl, benzotriazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzoxadiazolyl or benzothiadiazolyl ring,
wherein said R2 bicyclic ring is optionally mono- or di-substituted independently on carbon or nitrogen with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino substituents are optionally mono-substituted with hydroxy, (C1-C4)alkanoylamino, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to five fluorines;
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the Z Group of compounds designated the AA Group, contains those compounds wherein
R2 is a quinolinyl, isoquinolinyl, indazolyl or benzimidazolyl ring,
wherein said R2 bicyclic ring is optionally mono- or di-substituted independently with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl,
wherein said (C1-C4)alkoxy or (C1-C4)alkyl substituents are optionally mono-substituted with hydroxy, (C1-C4)alkanoylamino, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to five fluorines; or the pharmaceutically acceptable salts thereof.
Especially preferred compounds of Formula I are the compounds
[5-cyclopropyl-1-(quinolin-8-yl)-1H-pyrazole-4-carbonyl]guanidine; or
[5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carbonyl]guanidine;
or the pharmaceutically acceptable salts of said compounds.
Preferred salts of the immediately preceding compound are the mono -or di-mesylate salts.
Other especially preferred compounds of Formula I are the compounds
[1-(8-Bromoquinolin-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(6-Chloroquinolin-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(Indazol-7-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(Benzimidazol-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(1-Isoquinolyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[5-Cyclopropyl-1-(4-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
or the pharmaceutically acceptable salts of said compounds.
Especially preferred compounds within the AA Group are compounds wherein
a. R2 is 8-bromoquinolin-5-yl;
b. R2 is 6-chloroquinolin-5-yl;
c. R2 is indazol-7-yl;
d. R2 is benzimidazol-5-yl;
e. R2 is 1-isoquinolyl;
f. R2 is 4-quinolinyl;
or the pharmaceutically acceptable salts of said compounds.
A preferred group of compounds, designated the BB Group, contains those comounds having the Formula I as shown above wherein Z is 
R1 is (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R1 substituent optionally mono- or di-substituted independently with (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R2 is a five to six membered nonaromatic heterocyclic ring having one to two heteroatoms selected independently from nitrogen, sulfur and oxygen or R2 is unsubstituted (C1-C4)alkyl or unsubstituted (C3-C7)cycloalkyl; or R2 is phenyl(C1-C4)alkyl, or a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R2 substituents optionally substituted on carbon or nitrogen with up to three substituents independently selected from R6, R7 and R8, wherein one of R6, R7 and R8 is optionally a partially saturated, fully saturated, or fully unsaturated three to seven membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen optionally substituted with (C1-C4)alkyl and additionally R6, R7 and R8 are optionally hydroxy, nitro, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, formyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl, (C2-C4)alkenyl, (C2-C4)alkynyl or (C5-C7)cycloalkenyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino or (C3-C7)cycloalkyl R6, R7 and R8 substituents are optionally mono-substituted independently with hydroxy, (C1-C4)alkoxycarbonyl, (C3-C7)cycloalkyl, (C1-C4)alkanoyl, (C1-4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, nitro, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines,
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the BB Group of compounds designated the CC Group, contains those compounds wherein
R1 is (C1-C4)alkyl; and
R2 is a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently having one to three heteroatoms selected independently from nitrogen, sulfur and oxygen,
said R2 bicyclic ring is optionally mono-substituted on carbon or nitrogen with a fully saturated or fully unsaturated five to six membered ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen, said ring optionally mono-substituted with (C1-C4)alkyl
said R2 bicyclic ring is also optionally mono- or di-substituted independently on carbon or nitrogen with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino are optionally mono-substituted with hydroxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the CC Group of compounds designated the DD Group, contains those compounds wherein
R2 is a quinazolinyl, phthalazinyl, quinolinyl, isoquinolinyl, cinnolinyl, benzodioxanyl, quinoxalinyl, benzopyranyl, benzothiophenyl, benzodioxolyl, benzirnidazolyl, indazolyl, indolyl, benzotriazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzoxadiazolyl or benzothiadiazolyl ring,
wherein said R2 bicyclic ring is optionally mono- or di-substituted independently with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino substituents are optionally mono-substituted with hydroxy, (C1-C4)alkanoylamino, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to five fluorines;
or the pharmaceutically acceptable salts thereof.
Especially preferred compounds of Formula I are the compounds
[1-(Indazol-6-yl)-5-ethyl-1H-pyrazole-4carbonyl]guanidine;
[1-(Indazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(Benzimidazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(1-Methylbenzimidazol-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine
[1-(5-Quinolinyl)-5-n-propyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(5-Quinolinyl)-5-isopropyl-1H-pyrazole-4-carbonyl]guanidine;
[5-Ethyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-Methylbenzimidazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(1,4-Benzodioxan-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(Benzotriazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(3-Chloroindazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(5-Quinolinyl)-5-butyl-1H-pyrazole-4-carbonyl]guanidine;
[5-propyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[5-lsopropyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
or pharmaceutically acceptable salts of said compounds.
Especially preferred compounds within the DD Group are compounds wherein
a.
R1 is ethyl; and
R2 is indazol-6-yl;
b.
R1 is ethyl; and
R2 is indazol-5-yl;
c.
R1 is ethyl; and
R2 is benzimidazol-5-yl;
d.
R1 is ethyl; and
R2 is 1-methylbenzimidazol-6-yl;
e.
R1 is n-propyl; and
R2 is 5-quinolinyl;
f.
R1 is isopropyl; and
R2 is 5-quinolinyl;
g.
R1 is ethyl; and
R2 is 6-quinolinyl;
h.
R1 is ethyl; and
R2 is 2-methylbenzimidazol-5-yl;
i.
R1 is ethyl; and
R2 is 1,4-benzodioxan-6-yl;
j.
R1 is ethyl; and
R2 is benzotriazol-5-yl;
k.
R1 is ethyl; and
R2 is 3-Chloroindazol-5-yl;
l.
R1 is butyl; and
R2 is 5-quinolinyl;
m.
R1 is n-propyl; and
R2 is 6-quinolinyl;
n.
R1 is isopropyl; and
R2 is 6-quinolinyl;
or the pharmaceutically acceptable salts of said compounds.
A preferred group of compounds, designated the EE Group, contains those compounds having the Formula I as shown above wherein Z is 
R1 is (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R1 substituent optionally mono- or di-substituted independently with (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R2 is a five to six membered nonaromatic heterocyclic ring having one to two heteroatoms selected independently from nitrogen, sulfur and oxygen or R2 is unsubstituted (C1-C4)alkyl or unsubstituted (C3-C7)cycloalkyl; or R2 is phenyl(C1-C4)alkyl, or a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R2 substituents optionally substituted on carbon or nitrogen with up to three substituents independently selected from R6, R7 and R8, wherein one of R6, R7 and R8 is optionally a partially saturated, fully saturated, or fully unsaturated three to seven membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen optionally substituted with (C1-C4)alkyl and additionally R6, R7 and, R8 are optionally hydroxy, nitro, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, formyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl, (C2-C4)alkenyl, (C2-C4)alkynyl or (C5-C7)cycloalkenyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino or (C3-C7)cycloalkyl R6, R7 and R8 substituents are optionally mono-substituted independently with hydroxy, (C1-C4)alkoxycarbonyl, (C3-C7)cycloalkyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, nitro, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines,
or the, pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the EE Group of compounds designated the FF Group, contains those compounds wherein
R1 is (C1-C4)alkyl; and
R2 is a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently having one to three heteroatoms selected independently from nitrogen, sulfur and oxygen,
said R2 bicyclic ring is optionally mono-substituted on carbon or nitrogen with a fully saturated or fully unsaturated five to six membered ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen, said ring optionally mono-substituted with (C1-C4)alkyl
said R2 bicyclic ring is also optionally mono- or di-substituted independently on carbon or nitrogen with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino are optionally mono-substituted with hydroxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines.
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the FF Group of compounds designated the GG Group, contains those compounds wherein
R2 is a quinazolinyl, phthalazinyl, quinolinyl, isoquinolinyl, cinnolinyl, benzodioxanyl, quinoxalinyl, benzopyranyl, benzothiophenyl, benzodioxolyl, benzimidazolyl, indazolyl, indolyl, benzotriazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzoxadiazolyl or benzothiadiazolyl ring,
wherein said R2 bicyclic ring is optionally mono- or di-substituted independently with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N ,Nxe2x80x94(C1-C4)alkylaminosulfonyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino substituents are optionally mono-substituted with hydroxy, (C1-C4)alkanoylamino, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to five fluorines;
or the pharmaceutically acceptable salts thereof.
Especially preferred compounds of Formula I are the compounds
[1-(Indazol-7-yl)-3-methyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2,1,3-Benzothiadiazol-4-yl)-3-methyl-1H-pyrazole-4-carbonyl]guanidine;
[3-Methyl-1-(quinolin-5-yl)-1H-pyrazole-4-carbonyl]guanidine;
or the pharmaceutically acceptable salts of said compounds.
Especially preferred compounds within the GG Group of compounds are compounds wherein
a.
R1 is methyl; and
R2 is indazol-7-yl;
b.
R1 is methyl; and
R2 is 2,1,3-benzothiadiazol-4-yl;
c.
R1 is methyl; and
R2 is quinolin-5-yl;
or the pharmaceutically acceptable salts of said compounds.
A preferred group of compounds, designated the HH Group, contains those compounds having the Formula I as shown above wherein Z is 
R4 is (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R4 substituent optionally mono- or di-substituted independently with (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl or (C1-C4)alkylsulfonyl; and
R5 is a five to six membered nonaromatic heterocyclic ring having one to two heteroatoms selected independently from nitrogen, sulfur and oxygen or R5 is unsubstituted (C1-C4)alkyl or (C3-C7)cycloalkyl; or R5 is phenyl(C1-C4)alkyl, or a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen, said R5 substituents optionally substituted on carbon or nitrogen with; up to three substituents independently selected from R6, R7 and R8, wherein one of R6, R7 and R8 is optionally a partially saturated, fully saturated, or fully unsaturated three to seven membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen optionally substituted with (C1-C4)alkyl and additionally R6, R7 and R8 are optionally hydroxy, nitro, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, formyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl, (C2-C4)alkenyl, (C2-C4)alkynyl or (C5-C7)cycloalkenyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino or (C3-C7)cycloalkyl R6, R7 and R8 substituents are optionally mono-substituted independently with hydroxy, (C1-C4)alkoxycarbonyl, (C3-C7)cycloalkyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, nitro, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines,
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the HH Group of compounds designated the II Group, contains those compounds wherein
R4 is (C1-C4)alkyl; and
R5 is a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently having one to three heteroatoms selected independently from nitrogen, sulfur and oxygen,
said R5 bicyclic ring is optionally mono-substituted on carbon with a fully saturated or fully unsaturated five to six membered ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen, said ring optionally mono-substituted with (C1-C4)alkyl
said R5 bicyclic ring is also optionally mono- or di-substituted independently on carbon or nitrogen with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkysulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkythio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino are optionally mono-substituted with hydroxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C0-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines.
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the II Group of compounds designated the JJ Group, contains those compounds wherein
R5 is a quinazolinyl, phthalazinyl, quinolinyl, isoquinolinyl, cinnolinyl, benzodioxanyl, quinoxalinyl, benzopyranyl, benzothiophenyl, benzodioxolyl, benzitnidazolyl, indazolyl, indolyl, benzotriazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzoxadiazolyl or benzothiadiazolyl ring,
wherein said R5 bicyclic ring is optionally mono- or di-substituted independently with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino substituents are optionally mono-substituted with hydroxy, (C1-C4)alkanoylamino, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to five fluorines;
or the pharmaceutically acceptable salts thereof.
A preferred group of compounds, designated the KK Group, contains those compounds having the Formula I as shown above wherein Z is
Z is 
R2 is (C1-C4)alkyl, (C3-C7)cycloalkyl, M or M(C1-C4)alkyl, any of said previous (C1-C4)alkyl moieties optionally having from one to nine fluorines; said (C1-C4)alkyl or (C3-C4)cycloalkyl optionally mono- or di-substituted independently with hydroxy; (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, (C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl; and said (C3-C4)cycloalkyl optionally having from one to seven fluorines;
wherein M is a partially saturated, fully saturated or fully unsaturated five to eight membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen, or, a bicyclic ring consisting of two fused partially saturated, fully saturated or fully unsaturated three to six membered rings, taken independently, optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen;
said M is optionally substituted, on one ring if the moiety is monocyclic, or one or both rings if the moiety is bicyclic, on carbon or nitrogen with up to three substituents independently selected from R6, R7 and R8, wherein one of R6, R7 and R8 is optionally a partially saturated, fully saturated, or fully unsaturated three to seven membered ring optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen optionally substituted with (C1-C4)alkyl and additionally R6, R7 and R8 are optionally hydroxy, nitro, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, formyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl, (C2-C4)alkenyl, (C2-C4)alkynyl or (C5-C7)cycloalkenyl,
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino or (C3-C7)cycloalkyl R6, R7 and R8 substituents are optionally mono-substituted independently with hydroxy, (C1-C4)alkoxycarbonyl, (C3-C7)cycloalkyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, thiol, nitro, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines; and
R3 is (C1-C4)alkyl, (C3-C7)cycloalkyl, phenyl or phenyl(C1-C4)alkyl, said (C1-C4)alkyl optionally substituted with from one to nine fluorines, said R3 substituent optionally mono- or di-substituted independently with (C1-C4)alkoxy, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or (C1-C4)alkyl, or a pharmaceutically acceptable salt thereof.
A group of compounds which is preferred among the KK Group of compounds designated the LL Group, contains those compounds wherein
R3 is (C1-C4)alkyl;
R2 is phenyl, said phenyl optionally mono-substituted on carbon with a fully saturated or fully unsaturated five to six membered ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen, said ring optionally mono-substituted with (C1-C4)alkyl said R2 ring is also optionally mono- or di-substituted independently on carbon with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino are optionally mono-substituted with hydroxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines,
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the KK Group of compounds designated the MM Group, contains those compounds wherein
R3 is (C1-C4)alkyl;
R2 is a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently
said R2 bicyclic ring is optionally mono-substituted on carbon with a fully saturated or fully unsaturated five to six membered ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen, said ring optionally mono-substituted with (C1-C4)alkyl
said R2 bicyclic ring is also optionally mono- or di-substituted independently on carbon with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamirtosulfonyl
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino are optionally mono-substituted with hydroxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines,
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the KK Group of compounds designated the NN Group, contains those compounds wherein
R3 is (C1-C4)alkyl;
R2 is a five to six membered monocyclic aromatic ring having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen;
said R2 ring is optionally mono-substituted on carbon with a fully saturated or fully unsaturated five to six membered ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen, said ring optionally mono-substituted with (C1-C4)alkyl
said R2 ring is also optionally mono- or di-substituted independently on carbon or nitrogen with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino are optionally mono-substituted with hydroxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkyliminosulfonyl or optionally substituted with one to nine fluorines
or the pharmaceutically acceptable salts thereof.
A group of compounds which is preferred among the KK Group of compounds designated the OO Group, contains those compounds wherein
R3 is (C1-C4)alkyl;
R2 is a bicyclic ring consisting of two fused five and/or six membered partially saturated, fully saturated or fully unsaturated rings taken independently having one to three heteroatoms selected independently from nitrogen, sulfur and oxygen,
said R2 bicyclic ring is optionally mono-substituted on carbon or nitrogen with a fully saturated or fully unsaturated five to six membered ring optionally having one to two heteroatoms selected independently from oxygen, sulfur and nitrogen, said ring optionally mono-substituted with (C1-C4)alkyl
said R2 bicyclic ring is also optionally mono- or di-substituted independently on carbon or nitrogen with hydroxy, halo, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkyl, (C1-C4)alkanoyl, (C1-C4)alkanoyloxy, (C1-C4)alkanoylamino, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, cyano, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl
wherein said (C1-C4)alkoxy, (C1-C4)alkyl, (C1-C7)alkanoyl, (C1-C4)alkylthio, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino are optionally mono-substituted with hydroxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkanoylamino, (C1-C4)alkanoyloxy, (C1-C4)alkoxycarbonylamino, sulfonamido, (C1-C4)alkylsulfonamido, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkylthio, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylaminosulfonyl or optionally substituted with one to nine fluorines
or the pharmaceutically acceptable salts thereof.
Another aspect of this invention is directed to the esters of
5-cyclopropyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carboxylate,
5-methyl-1-(6-quinolinyl)-1H-pyrazole-4-carboxylate,
5-methyl-1-naphthalenyl-1H-pyrazole-4-carboxylate,
5-cyclopropyl-1-(quinolin-8-yl)-1H-pyrazole-4-carboxylate,
5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carboxylate,
5-ethyl-1-(quinolin-5-yl)-1H-pyrazole-4-carboxylate or
n-butyl 1-(isoquinolin-5-yl)-3-methyl-1H-pyrazole-4-carboxylate wherein said esters are benzyl, (C1-C6)alkyl or (C4-C8)cycloalkyl, said (C4-C8)cycloalkyl optionally mono-substituted with (C1-C4)alkyl
or a salt of said esters.
Yet another aspect of this invention is directed to the following compounds
5-methyl-2-(5-quinolinyl)-2H-1,2,3-triazole-4-carboxylic acid,
5-methyl-2-(5-isoquinolinyl)-2H-1,2,3-triazole-4-carboxylic acid,
2-(1-nabphthalenyl)-5-methyl-2H-1,2,3-triazole-4-carboxylic acid,
5-methyl-1-(6-quinolinyl)-1H-pyrazole-4-carboxylic acid,
5-methyl-1-naphthalenyl-1H-pyrazole-4-carboxylic acid,
5-cyclopropyl-1-(quinolin-8-yl)-1H-pyrazole-4-carboxylic acid,
5-cyclopropyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carboxylic acid,
5-ethyl-1-(quinolin-5-yl)-1H-pyrazole-4-carboxylic acid,
5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carboxylic acid or
1-(isoquinolin-5-yl)-3-methyl-1H-pyrazole-4-carboxylic acid or the acid chlorides thereof or a salt of said compounds or of said acid chlorides.
Another aspect of this invention is a method of treating a mammal (e.g., human) having a disease or condition mediated by NHE-1 by administering a pharmaceutically acceptable amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug to the mammal.
Another aspect of this invention is directed to a method of reducing tissue damage (e.g., substantially preventing tissue damage, inducing tissue protection) resulting from ischemia comprising administering to a mammal (e.g., a female or male human) in need of such treatment a therapeutically effective amount of a compound of Formula I a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Preferred ischemic tissues taken individually or as a group are wherein the ischemic tissue is cardiac, brain, liver, kidney, lung, gut, skeletal muscle, spleen, pancreas, nerve, spinal cord, retina tissue, the vasculature, or intestinal tissue.
An especially preferred ischemic tissue is cardiac tissue.
It is especially preferred that the compounds are administered to prevent perioperative myocardial ischemic injury.
Preferably, the compounds of this invention are administered prophylactically.
The ischemic damage may occur during organ transplantation.
Preferably, the compounds of this invention are administered prior to, during or shortly after, cardiac surgery or non-Cardiac surgery.
In one aspect of this invention a compound of Formula I is administered locally.
A preferred dosage is about 0.001 to 100 mg/kg/day of the Formula I compound a prodrug thereof. An especially preferred dosage is about 0.01 to 50 mg/kg/day of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method of reducing myocardial tissue damage (e.g., substantially preventing tissue damage, inducing tissue protection) during surgery (e.g., coronary artery bypass grafting (CABG) surgeries, vascular surgeries, percutaneous transluminal coronary angioplasty (PTCA) or any percutaneous transluminal coronary intervention (PTC1), organ transplantation, or other non-Cardiac surgeries) comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method of reducing myocardial tissue damage (e.g., substantially preventing tissue damage, inducing tissue protection) in patients presenting with ongoing cardiac (acute coronary syndromes, e.g. myocardial infarction or unstable angina) or cerebral ischemic events (e.g. stroke) comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a chronic method of reducing myocardial tissue damage (e.g., substantially preventing tissue damage, inducing tissue protection) in a patient with diagnosed coronary heart disease (e.g. previous myocardial infarction or unstable angina) or patients who are at high risk for myocardial infarction (age  greater than 65 and two or more risk factors for coronary heart disease) comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method of preventing ischemic damage comprising the chronic oral administration to a mammal in need of such treatment of a therapeutically effective amount of a compound of Formula I a prodrug of said compound or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating cardiovascular diseases comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating arteriosclerosis comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating hypertension comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating arrhythmia comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating angina pectoris comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating cardiac hypertrophy comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating renal diseases comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating diabetic complications comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating restenosis comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating diseases of cell proliferation comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating cancerous diseases comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating fibrotic diseases comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating glomerular nephrosclerosis comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating organ hypertrophies or hyperplasias comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating pulmonary fibrosis comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating cerebro ischemic disorders comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating myocardial stunning comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating myocardial dysfunction comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating cerebrovascular diseases comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating organ hypertrophies or hyperplasias comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Another aspect of this invention is directed to a method for treating organ hypertrophies or hyperplasias comprising administering to a mammal (e.g., a female or male human) a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
This invention is also directed to pharmaceutical compositions which comprise a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug and a pharmaceutically acceptable carrier.
This invention is also directed to pharmaceutical compositions for the reduction of tissue damage resulting from ischemia which comprise a therapeutically effective amount of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug and a pharmaceutically acceptable carrier.
Yet another aspect of this invention are combinations of a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug and other compounds as described below.
Yet another aspect of this invention is directed to pharmaceutical compositions comprising a compound of Formula I or a pharmaceutically acceptable salt or prodrug thereof and a cardiovascular agent and for the use of such compositions for the reduction of tissue damage resulting from tissue ischemia in mammals (e.g., humans, male or female).
In the above pharmaceutical compositions and methods preferred Formula I compounds include the preferred groups of compounds described above labeled as Group A-to Group OO.
Another aspect of this invention is a method of reducing tissue damage (e.g., substantially preventing tissue damage, inducing tissue protection) resulting from or which could result from ischemia comprising administering to a mammal (e.g., a female or male human)
a. a first compound, said first compound being a Formula I compound, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug; and
b. a second compound, said second compound being a cardiovascular agent
wherein the amounts of the first and second compounds result in a therapeutic effect.
Another aspect of this invention is a kit comprising:
a. a therapeutically effective amount of a Formula I compound, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug and a pharmaceutically acceptable carrier, vehicle or diluent in a first unit dosage form;
b. a therapeutically effective amount of a cardiovascular agent and a pharmaceutically acceptable carrier, vehicle or diluent in a second unit dosage form; and
c. means for containing said first and second dosage forms wherein the amounts of the first and second compounds result in a therapeutic effect.
In the above combination compositions, combination methods and kits, preferably the cardiovascular agents are for example, xcex2-blockers (e.g., acebutolol, atenolol, bopindolol, labetolol, mepindolol, nadolol, oxprenol, pindolol, propranolol, sotalol), calcium channel blockers (e.g., amlodipine, nifedipine, nisoldipine, nitrendipine, verapamil), potassium channel openers, adenosine, adenosine agonists, ACE inhibitors (e.g., captopril, enalapril), nitrates (e.g., isosorbide dinitrate, isosorbide 5-mononitrate, glyceryl trinitrate), diuretics (e.g., hydrochlorothiazide, indapamide, piretanide, xipamide), glycosides (e.g., digoxin, metildigoxin), thrombolytics (e.g. tPA), platelet inhibitors (e.g., reopro), aspirin, dipyridamol, potassium chloride, clonidine, prazosin or adenosine A3 receptor agonists.
In the above combination compositions, combination methods and kits preferred Formula I compounds include the preferred groups of compounds described above labeled as Group A to Group OO.
This invention is also directed to a pharmaceutical combination composition comprising: a therapeutically effective amount of a composition comprising
a first compound, said first compound being a Formula I compound, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug;
a second compound, said second compound being a glycogen phosphorylase inhibitor; and/or optionally
a pharmaceutical carrier, vehicle or diluent.
Another aspect of this invention is a method of reducing tissue damage (e.g., substantially preventing tissue damage, inducing tissue protection) resulting from or which could result from ischemia comprising administering to a mammal (e.g., a female or male human)
a. a first compound, said first compound being a Formula I compound, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug; and
b. a second compound, said second compound being a glycogen phosphorylase inhibitor wherein the amounts of the first and second compounds result in a therapeutic effect.
Another aspect of this invention is a kit comprising:
a. a therapeutically effective amount of a Formula I compound, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug and a pharmaceutically acceptable carrier, vehicle or diluent in a first unit dosage form;
b. a therapeutically effective amount of a glycogen phosphorylase inhibitor and a pharmaceutically acceptable carrier, vehicle or diluent in a second unit dosage form; and
c. means for containing said first and second dosage forms wherein the amounts of the first and second compounds result in a therapeutic effect.
In the above combination compositions, combination methods and kits preferred Formula I compounds include the preferred groups of compounds described above labeled as Group A to Group OO.
In the above combination compositions, combination methods and kits preferred glycogen phosphorylase inhibitors are
5-chloro-1H-indole-2-carboxylic acid [(1S)-((R)-hydroxy-dimethylcarbamoyl-methyl)-2-phenyl-ethyl]-amide,
5,6-dichloro-1H-indole-2-carboxylic acid {(1S)-[(R)-hydroxy-(methoxy-methyl-carbamoyl)-methyl]-2-phenyl-ethyl}-amide,
5-chloro-1H-indole-2-carboxylic acid {(1S)-[(R)-hydroxy-(methoxy-methyl-carbamoyl)-methyl]-2-phenyl-ethyl}-amide,
5-chloro-1H-indole-2-carboxylic acid ((1S)-{(R)-hydroxy-[(2-hydroxy-ethyl)-methyl-carbamoyl]-methyl}-2-phenyl-ethyl)-amide,
5-chloro-1H-indole-2-carboxylic acid {(1S)-[(R)-hydroxy-(methyl-pyridin-2-yl-carbamoyl)-methyl]-2-phenyl-ethyl}-amide or
5-chloro-1H-indole-2-carboxylic acid ((1S)-{(R)-hydroxy-[methyl-(2-pyridin-2-yl-ethyl)-carbamoyl]-methyl}-2-phenyl-ethyl)-amide.
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-(2R)-hydroxy-3-(4-methyl-piperazin-1-yl)-3-oxo-propyl]-amide hydrochloride,
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-(2R)-hydroxy-3-(3-hydroxy-azetidin-1-yl)-3-oxo-propyl]-amide,
5-chloro-1H-indole-2-carboxylic acid ((1S)-benzyl-(2R)-hydroxy-3-isoxazolidin-2-yl-3-oxo-propyl)-amide,
5-Chloro-1H-indole-2-carboxylic acid ((1S)-benzyl-(2R)-hydroxy-3-[1,2]oxazinan-2-yl-3-oxo-propyl)-amide,
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-(2R)-hydroxy-3-((3S)-hydroxy-pyrrolidin-1-yl)-3-oxo-propyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-3-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-(2R)-hydroxy-3-oxo-propyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-3-((3R,4S)-dihydroxy-pyrrolidin-1-yl)-(2R)-hydroxy-3-oxo-propyl]-amide or
5-chloro-1H-indole-2-carboxylic acid ((1S)-benzyl-(2R)-hydroxy-3-morpholin-4-yl-3-oxo-propyl)-amide.
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-(3-hydroxyimino-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [2-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [2-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [2-(1,1-dioxo-thiazolidin-3-yl)-2-oxo-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid (2-oxo-2-thiazolidin-3-yl-ethyl)-amide,
5-chloro-1H-indole-2-carboxylic acid [(1S)-(4-fluoro-benzyl)-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-((3RS)-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [2-oxo-2-((1RS)-oxo-1-thiazolidin-3-yl)-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [(1S)-(2-fluoro-benzyl)-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-(3-hydroxy-azetidin-1-yl)-2-oxo-ethyl]-amide,
5-Chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-(3-hydroxyimino-azetidin-1-yl)-2-oxo-ethyl]-amide or
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-(4-hydroxyimino-piperidin-1-yl)-2-oxo-ethyl]-amide.
This invention is also directed to a pharmaceutical combination composition comprising: a therapeutically effective amount of a composition comprising
a first compound, said first compound being a Formula I compound, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug;
a second compound, said second compound being an aldose reductase inhibitor; and/or optionally
a pharmaceutical carrier, vehicle or diluent.
Another aspect of this invention is a method of reducing tissue damage (e.g., substantially preventing tissue damage, inducing tissue protection) resulting from or which could result from ischemia comprising administering to a mammal (e.g., a female or male human)
a. a first compound, said first compound being a Formula I compound, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug; and
b. a second compound, said second compound being an aldose reductase inhibitor
wherein the amounts of the first and second compounds result in a therapeutic effect.
Another aspect of this invention is a kit comprising:
a. a therapeutically effective amount of a Formula I compound, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug and a pharmaceutically acceptable carrier, vehicle or diluent in a first unit dosage form;
b. a therapeutically effective amount of an aldose reductase inhibitor and a pharmaceutically acceptable carrier, vehicle or diluent in a second unit dosage form; and
c. means for containing said first and second dosage forms wherein the amounts of the first and second compounds result in a therapeutic effect.
In the above combination compositions, combination methods and kits preferred Formula I compounds include the preferred groups of compounds described above labeled as Group A to Group OO.
In the above combination compositions, combination methods and kits a preferred aldose reductase inhibitor is zopolrestat: 1-phthalazineacetic acid, 3,4-dihydro-4-oxo-3-[[5-trifluoromethyl)-2-benzothiazolyl]methyl]-.
In the methods of treatment as applied to the combinations described above the following are preferred administration routes, modes etc.
Preferred ischemic tissues taken individually or as a group are wherein the ischemic tissue is cardiac, brain, liver, kidney, lung, gut, skeletal muscle, spleen, pancreas, nerve, spinal cord, retina tissue, the vasculature, or intestinal tissue.
An especially preferred ischemic tissue is cardiac tissue.
It is especially preferred that the compounds are administered to prevent perioperative myocardial ischemic injury.
Preferably, the compounds of this invention are administered prophylactically.
The ischemic damage may occur during organ transplantation.
Preferably, the compounds of this invention are administered prior to, during or shortly after, cardiac surgery or non-cardiac surgery.
In one aspect of this invention the compounds are administered locally.
In one aspect of this method myocardial tissue damage is reduced during surgery.
In another aspect of this method myocardial tissue damage is reduced in patients presenting with ongoing cardiac or cerebral ischemic events.
In yet another aspect of this method myocardial tissue damage is reduced by chronic administration of the combination in a patient with diagnosed coronary heart disease.
The term xe2x80x9creductionxe2x80x9d is intended to include partial prevention or prevention which, although greater than that which would result from taking no compound or from taking a placebo, is less than 100% in addition to substantially total prevention.
The term xe2x80x9cdamage resulting from ischemiaxe2x80x9d as employed herein refers to conditions directly associated with reduced blood flow to tissue, for example due to a clot or obstruction of blood vessels which supply blood to the subject tissue and which result, inter alia, in lowered oxygen transport to such tissue, impaired tissue performance, tissue dysfunction and/or necrosis. Alternatively, where blood flow or organ perfusion may be quantitatively adequate, the oxygen carrying capacity of the blood or organ perfusion medium may be reduced, e.g., in hypoxic environment, such that oxygen supply to the tissue is lowered, and impaired tissue performance, tissue dysfunction, and/or tissue necrosis ensues.
The term xe2x80x9ctreatingxe2x80x9d, xe2x80x9ctreatxe2x80x9d or xe2x80x9ctreatmentxe2x80x9d as used herein includes preventative (e.g., prophylactic) and palliative treatment.
By xe2x80x9cpharmaceutically acceptablexe2x80x9d it is meant the carrier, diluent, excipients, and/or salt must be compatible with the other ingredients of the formulation, and not deleterious to the recipient thereof.
The expression xe2x80x9cprodrugxe2x80x9d refers to compounds that are drug precursors which following administration, release the drug in vivo via some chemical or physiological process (e.g., a prodrug on being brought to the physiological pH or through enzyme action is converted to the desired drug form).
Exemplary five to six membered aromatic rings optionally having one or two heteroatoms selected independently from oxygen, nitrogen and sulfur are phenyl, furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, pyridinyl, pyridiazinyl, pyrimidinyl and pyrazinyl.
Exemplary partially saturated, fully saturated or fully unsaturated five to eight membered rings optionally having one to three heteroatoms selected independently from oxygen, sulfur and nitrogen are cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and phenyl. Further exemplary five membered rings are furyl, thienyl, pyrrolyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrrolidinyl, 1,3-dioxolanyl, oxazolyl, thiazolyl, imidazolyl, 2H-imidazolyl 2-imidazolinyl, imidazolidinyl, pyrazolyl, 2-pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, 1,2-dithiolyl, 1,3-dithiolyl, 3H-1,2-oxathiolyl, 1,2,3-oxadizaolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-trizaolyl, 1,3,4-thiadiazolyl, 3H-1,2,3-dioxazolyl, 1,2,4-dioxazolyl, 1,3,2-dioxazolyl, 1,3,4-dioxazolyl, 5H-1,2,5-oxathiazolyl and 1,3-oxathiolyl.
Further exemplary six membered rings are 2H-pyranyl, 4H-pyranyl, pyridinyl, piperidinyl, 1,2-dioxinyl, 1,3-dioxinyl, 1,4-dioxanyl, morpholinyl, 1,4-dithianyl, thiomoirpholinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, 1,3,5-triazinyl, 1,2,4-triazinyl, 1,2,3-triazinyl, 1,3,5-trithianyl, 4H-1,2-oxazinyl, 2H-1,3-oxazinyl, 6H-1,3-oxazinyl, 6H-1,2-oxazinyl, 1,4-oxazinyl, 2H-1,2-oxazinyl, 4H-1,4-oxazinyl, 1,2,5-oxathiazinyl, 1,4-oxazinyl, o-isoxazinyl, p-isoxazinyl, 1,2,5-oxathiazinyl, 1,2,6-oxathiazinyl and 1,4,2-oxadiazinyl.
Further exemplary seven membered rings are azepinyl, oxepinyl, thiepinyl and 1,2,4-diazepinyl.
Further exemplary eight membered rings are cyclooctyl, cyclooctenyl and cyclooctadienyl.
Exemplary bicyclic rings consisting of two fused partially saturated, fully saturated or fully unsaturated five and/or six membered rings, taken independently, optionally having one to four heteroatoms selected independently from nitrogen, sulfur and oxygen are indolizinyl, indolyl, isoindolyl, indolinyl, cyclopenta(b)pyridinyl, pyrano(3,4-b)pyrrolyl, benzofuryl, isobenzofuryl, benzo(b)thienyl, benzo(c)thienyl, 1H-indazolyl, indoxazinyl, benzoxazolyl, anthranilyl, benzimidazolyl, benzthiazolyl, purinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, indenyl, isoindenyl, naphthyl, tetralinyl, decalinyl, 2H-1-benzopyranyl, pyrido(3,4-b)-pyridinyl, pyrido(3,2-b)-pyridinyl, pyrido(4,3-b)-pyridinyl, 2H-1,3-benzoxazinyl, 2H-1,4-benzoxazinyl, 1H-2,3-benzoxazinyl, 4H-3,1-benzoxazinyl, 2H-1,2-benzoxazinyl and 4H-1,4-benzoxazinyl.
By alkylene is meant saturated hydrocarbon (straight chain or branched) wherein a hydrogen atom is removed from each of the terminal carbons. Exemplary of such groups (assuming the designated length encompases the particular example) are methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene).
By halo is meant chloro, bromo, iodo, or fluoro.
By alkyl is meant straight chain saturated hydrocarbon or branched saturated hydrocarbon. Exemplary of such alkyl groups (assuming the designated length encompasses the particular example) are methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tertiary butyl, pentyl, isopentyl, neopentyl, tertiary pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, hexyl, isohexyl, heptyl and octyl.
By alkoxy is meant straight chain saturated alkyl or branched saturated alkyl bonded through an oxygen. Exemplary of such alkoxy groups (assuming the designated length encompasses the particular example) are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tertiary butoxy, pentoxy, isopentoxy, neopentoxy, tertiary pentoxy, hexoxy, isohexoxy, heptoxy and octoxy .
As used herein the term mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-Cx)alkyl . . . refers to the (C1-Cx)alkyl moiety taken independently when it is di-N,Nxe2x80x94(C1-Cx)alkyl . . . (x refers to integers).
It is to be understood that if a carbocyclic or heterocyclic moiety may be bonded or otherwise attached to a designated substrate through differing ring atoms without denoting a specific point of attachment, then all possible points are intended, whether through a carbon atom or, for example, a trivalent nitrogen atom. For example, the term xe2x80x9cpyridylxe2x80x9d means 2-, 3-, or 4-pyridyl, the term xe2x80x9cthienylxe2x80x9d means 2-, or 3-thienyl, and so forth.
The expression xe2x80x9cpharmaceutically-acceptable saltxe2x80x9d refers to nontoxic anionic salts containing anions such as (but not limited to) chloride, bromide, iodide, sulfate, bisulfate, phosphate, acetate, maleate, fumarate, oxalate, lactate, tartrate, citrate, gluconate, methanesulfonate and 4-toluene-sulfonate. Where more than one basic moiety exists the expression includes multiple salts (e.g., di-salt). The expression also refers to nontoxic cationic salts such as (but not limited to) sodium, potassium, calcium, magnesium, ammonium or protonated benzathine (N,Nxe2x80x2-dibenzylethylenediamine), choline, ethanolamine, diethanolamine, ethylenediamine, meglamine (N-methyl-glucamine), benethamine (N-benzylphenethylamine), piperazine or tromethamine (2-amino-2-hydroxymethyl-1,3-propanediol).
As used herein, the expressions xe2x80x9creaction-inert solventxe2x80x9d and xe2x80x9cinert solventxe2x80x9d refers to a solvent or mixture of solvents which does not interact with starting materials, reagents, intermediates or products in a manner which adversely affects the yield of the desired product.
The chemist of ordinary skill will recognize that certain compounds of this invention will contain one or more atoms which may be in a particular stereochemical or geometric configuration, giving rise to stereoisomers and configurational isomers. All such isomers and mixtures thereof are included in this invention. Hydrates of the compounds of this invention are also included.
DMF means N,N-dimethylformamide. DMSO means dimethyl sulfoxide. THF means tetrahydrofuran.
The subject invention also includes isotopically-labelled compounds, which are identical to those recited in Formula I, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine, such as 2H, 3H, 13C, 14C, 15N, 18O, 17O, 31P, 32P, 35S, 18F, and 36Cl, respectively. Compounds of the present invention, prodrugs thereof, and pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labelled compounds of the present invention, for example those into which radioactive isotopes such as 3H and 14C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., 3H, and carbon-14, i.e., 14C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium, i.e., 2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labelled compounds of Formula I of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples below, by substituting a readily available isotopically labelled reagent for a non-isotopically labelled reagent.
Other features and advantages will be apparent from the specification and claims which describe the invention.
In general the compounds of this invention can be made by processes which include processes known in the chemical arts, particularly in light of the description contained herein. Certain processes for the manufacture of the compounds of this invention are provided as further features of the invention and are illustrated by the following reaction schemes. Other processes are described in the experimental section.
Briefly, in general, a compound of the Fomula Zxe2x80x94C(O)OH is coupled with guanidine in the presence of a suitable coupling agent. 
According to Scheme I the Formula IA compound, wherein R4 is as described above, is dissolved or suspended in an aqueous alkali metal hydroxide solution (e.g. 1 N sodium hydroxide) along with sodium nitrite and the mixture is added to an aqueous acidic solution (e.g. 10% v/v sulfuric acid) at a pH of about 0 at a temperature of about 0xc2x0 C. to about 5xc2x0 C. for about 30 min to about 1 hour. The resulting mixture is filtered to yield the Formula II oxime. Alternatively, the Formula IA compound is dissolved in 1:1 acetic acid/propionic acid and solid sodium nitrite is added at about 0xc2x0 C. The reaction mixture is stirred at about 0xc2x0 C. for about 2 hours, then poured into ice water and the Formula II oxime is obtained by filtration.
The Formula II compound is reacted with a Formula III compound, wherein R5 is as described above in a protic solvent such as ethanol at a temperature of about 50xc2x0 C. to about 110xc2x0 C. for about 10 min to about 1 hour to form the Formula IV hydrazone.
The Formula IV hydrazone is cyclized and hydrolyzed to the Formula V triazole in an alcoholic solvent such as 2-ethoxyethanol under basic conditions (e.g., potassium hydroxide) at a temperature of about 100xc2x0 C. to about 175xc2x0 C. for about xc2xd hour to about 2 hours followed by acidification to yield the Formula V triazole acid.
The Formula V acid is coupled with guanidine in the presence of a suitable coupling agent. A suitable coupling agent is one which transforms a carboxylic acid into a reactive species which forms an amide linkage on reaction with an amine.
The coupling agent may be a reagent which effects this condensation in a one pot process when mixed together with the carboxylic acid and guanidine. Exemplary coupling reagents are 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-hydroxybenzotriazole (EDC/HBT), dicydohexylcarbodiimide/hydroxybenzotriazole(HBT), 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), and diethylphosphorylcyanide. The coupling is performed in an inert solvent, preferably an aprotic solvent at a temperature of about xe2x88x9220xc2x0 C. to about 50xc2x0 C. for about 1 to about 48 hours, in the presence of excess guanidine as base. Exemplary solvents include acetonitrile, dichloromethane, dimethylformamide and chloroform or mixtures thereof.
The coupling agent may also be that agent which converts the carboxylic acid to an activated intermediate which is isolated and/or formed in a first step and allowed to react with guanidine in a second step. Examples of such coupling agents and activated intermediates are thionyl chloride or oxalyl chloride to form the acid chloride, cyanuric fluoride to form an acid fluoride or an alkyl chloroformate such as isobutyl or isopropenyl chloroformate or propanephosphonic anhydride (propanephosphonic acid anhydride, PPA) (with a tertiary amine base) to form a mixed anhydride of the carboxylic acid, or carbonyldiimidazole to form an acylimidazole. If the coupling agent is oxalyl chloride, it is advantageous to employ a small amount of dimethylformamide as cosolvent with another solvent (such as dichloromethane) to catalyze the formation of the acid chloride. This activated acid derivative may be coupled by mixing with excess guanidine in an appropriate solvent together with an appropriate base. Appropriate solvent/base combinations are for example, dichloromethane, dimethylformamide or acetonitrile or mixtures thereof in the presence of excess guanidine as base. Other appropriate solvent/base combinations include water or a ((C1-C5)alcohol) or a mixture thereof together with a cosolvent such as dichloromethane, tetrahydrofuran or dioxane and a base such as sodium, potassium or lithium hydroxide in sufficient quantity to consume the acid liberated in the reaction. Use of these coupling agents and appropriate selection of solvents and temperatures are known to those skilled in the art or can be readily determined from the literature. These and other exemplary conditions useful for coupling carboxylic acids are described in Houben-Weyl, Vol XV, part II, E. Wunsch, Ed., G. Theime Verlag, 1974, Stuttgart; M. Bodansky, Principles of Peptide Synthesis, Springer-Verlag, Berlin 1984; and The Peptides, Analysis, Synthesis and Biology (ed. E. Gross and J. Meienhofer), vols 1-5 (Academic Press, NY 1979-1983).
According to Scheme II, the Formula X primary amine wherein R5 is as described above is reacted with a Formula XI xcex1-diazo-xcex2-keto-ester wherein R4 is as described above, and R is lower alkyl, in the presence of titanium tetrachloride analogously to the method described in Eguchi S. et al. Synthesis 1993, 793 to form the Formula XII triazole carboxylic acid ester. The Formula XII ester is converted directly to the acylguanidine XIII by reaction with guanidine in an alcoholic solvent at a temperature of about 60 to about 110xc2x0 C., preferably refluxing methanol, for a period of 8 to 20 hours.
According to Scheme III, the Formula XV compound wherein R4 and R5 are as described above is treated with Lawesson""s reagent (i.e., 2,4-bis(4-methoxyphenyl)-1,3-dithia-2,4-diphosphetane-2,4-disulfide) in an aprotic solvent such as dimethoxyethane at a temperature of about 20xc2x0 C. to about 120xc2x0 C. for about one to eight hours. The resulting thioamide is treated with an alkylating agent such as methyl iodide in a polar, inert solvent such as acetone, conveniently at ambient temperature for about eight hours to about forty-eight hours. The resulting compound is reacted with anhydrous hydrazine in an alcoholic solvent at a temperature of about 0xc2x0 C. to about 25xc2x0 C. for about one to eight hours to provide the Formula XVI compound (analogously as described in Doyle and Kurzer, Synthesis 1974, 583).
The Formula XVI compound is treated with a monoalkyloxalyl chloride in an aprotic solvent at a temperature of about 25xc2x0 C. to about 50xc2x0 C. for about one to eight hours to provide the Formula XVII carboxylic ester compound wherein R is lower alkyl. The Formula XVII ester is directly coupled with guanidine in an alcoholic solvent at a temperature of about 60xc2x0 C. to about 110xc2x0 C., preferably refluxing methanol, for a period of eight to twenty hours, to prepare the Formula XVIII triazole carbonyl guanidines.
According to Scheme IV the Formula XX compound wherein R5 is as described above is treated with methyl iodide in an inert solvent, conveniently at ambient temperature for about four to twenty-four hours. The resulting compound is reacted with anhydrous R4-hydrazine (wherein R4 is as described above) in an alcohoic solvent at a temperature of about 0xc2x0 C. to about 25xc2x0 C. for about one to eight hours to provide the Formula XXI amidrazone compound (analogously as described in Doyle and Kurzer, Synthesis 1974, 583).
The Formula XXI compound is treated with a monoalkyloxalyl chloride in an aprotic. solvent at a temperature of about 25xc2x0 C. to about 50xc2x0 C. for about one to eight hours to provide the Formula XXII carboxylic ester compound wherein R is lower alkyl. The Formula XXII ester is directly coupled with guanidine in an alcoholic solvent at a temperature of about 60xc2x0 C. to about 110xc2x0 C., preferably refluxing methanol, for a period of eight to twenty hours to prepare the Formula XXIII triazole carbonyl guanidines.
According to Scheme V the Formula XXV compound wherein R1 is as described above is combined with excess (CH3O)2C(R3)N(CH3)2 (N,N-dimethyl amide dimethyl acetal) wherein R3 is as described above, optionally in the presence of an acid catalyst such as p-toluenesulfonic acid at a temperature of about 90xc2x0 C. to about 110xc2x0 C. for about one to about two hours to prepare the Formula XXVI compound above.
The Formula XXVI compound is cyclized with a Formula XXVII compound, wherein R2 is as described above, in an inert solvent such as ethanol at a temperature of about 20xc2x0 C. to about 30xc2x0 C. for about 5 minutes to about one hour followed by heating to a temperature of about 70xc2x0 C. to about 110xc2x0 C. for about two hours to about four hours to form the Formula XXVIII pyrazole.
Alternatively, according to Scheme V the Formula XXV compound, wherein R1 is as described above, is combined with a triethylorthoester (i.e., R3C(OEt)3 wherein R3 is as described above) and acetic anhydride at a temperature of about 120xc2x0 C. to about 150xc2x0 C. for about two to about five hours to prepare the Formula XXXI compound.
The Formula XXXI compound is cyclized with a Formula XXVII compound, wherein R2 is as described above, to form the Formula XXVIII pyrazole.
The Formula XXVIII pyrazole is hydrolyzed with a base such as sodium hydroxide or lithium hydroxide in a solvent such as water and/or methanol and/or THF conveniently at ambient temperature or at elevated temperature (e.g., reflux) for about one hour to about five hours to prepare the Formula XXIX acid.
The Formula XXIX acid is coupled with guanidine in the presence of a suitable coupling agent as described for the above coupling of the Formula V acid and guanidine. In one embodiment, the Formula XXIX acid is activated with thionyl chloride at a temperature of about 60xc2x0 C. to about 90xc2x0 C. for about fifteen minutes to about two hours. The resulting activated acid chloride is combined with guanidine hydrochloride and an inorganic base (e.g., sodium hydroxide) in anhydrous tetrahydrofuran and optionally methanol and/or water. The solution is heated, conveniently at reflux, for about one hour to about eight hours to prepare the Formula XXX compound.
Alternatively according to Scheme V the Formula XXVIII compound can be directly converted to the Formula XXX compound by several methods. For example, the Formula XXVIII compound can be heated in the presence of excess guanidine, in a polar protic solvent for example, methanol or isopropanol at a suitable temperature conveniently, at reflux for about one to about seventy-two hours. This transformation may also be performed by repeatedly removing the solvent, for example removing ethanol or toluene about four times, from a mixture of the Formula XXVIII compound and excess guanidine at a pressure of about one to about 100 mmHg and at a temperature of about 25xc2x0 C. to about 95xc2x0 C. This reaction may also be performed in the absence of solvent by heating the mixture of the Formula XXVIII compound and excess guanidine at a temperature of about 100xc2x0 C. to about 180xc2x0 C., optionally at about a pressure of about 1 to about 100 mmHg for about five minutes to about eight hours.
According to Scheme VI, the Formula XXXV compound, wherein R3 is as described above, is reacted with the Formula XXXVI compound, wherein R1 and R2 are as described above, in an aprotic solvent at a temperature of about 0xc2x0 C. to about 25xc2x0 C. for about two hours to about twenty-four hours in the presence of an appropriate amine base, such as triethylamine, to form the Formula XXXVII compound.
The resulting Formula XXXVII compound is hydrolyzed and coupled with guanidine using one of the methods described in earlier Schemes, such as the method employing carbonyidiimidazole, to form the Formula XXXVIII compound.
According to Scheme VII, the Formula XL hydrazine, wherein R2 is as described above, is reacted with the appropriate Formula XLI compound to form the Formula XLII pyrazole ester wherein R is lower alkyl according to the method of Bajnati, A. and Hubert-Habart, M. Bull. Soc. Chim. France 1988, 540. The resulting pyrazole ester is converted to the Formula XLIII acyl guanidine using the hydrolysis and coupling methods described above.
According to Scheme VIII, the Formula L compound wherein R2 and R1 are as described above is transformed to the Formula LI lithium salt where R is lower alkyl according to the method described in J. Het. Chem. 1989, 26, 1389. The Formula LI lithium salt is combined with the Formula LII hydrazine, wherein R3 is as described above, in an inert solvent such as ethanol, in the presence of a mineral acid, at a temperature of about 20xc2x0 C. to about 30xc2x0 C. for about five minutes to about one hour followed by heating to a temperature of about 70xc2x0 C. to about 110xc2x0 C. for two hours to about four hours to form both the Formula LIII and LIV pyrazoles. The Formula LIII and LIV pyrazoles are converted to the Formula LV and LVI acyl guanidines respectively using the hydrolysis and coupling methods described above.
Some of the methods useful for the preparation of the compounds described herein may require protection of remote functionality (e.g., primary amine, secondary amine, carboxyl in Formula I precursors). The need for such protection will vary depending on the nature of the remote functionality and the conditions of the preparation methods. The need for such protection is readily determined by one skilled in the art. The use of such protection/deprotection methods is also within the skill in the art. For a general description of protecting groups and their use, see T. W. Greener, Protective Groups in Organic Synthesis, John Wiley and Sons, New York, 1991.
The starting materials and reagents for the above described compounds, are also readily available or can be easily synthesized by those skilled in the art using conventional methods of organic synthesis. For example, the aromatic hydrazines used in this invention can be prepared from the corresponding aromatic amines by diazotization followed by reduction conveniently using stannous chloride using procedures known to those skilled in the art. For example, many of the compounds used herein are related to, or are derived from compounds found in nature, in which there is a large scientific interest and commercial need, and accordingly many such compounds are commercially available or are reported in the literature or are easily prepared from other commonly available substances by methods which are reported in the literature.
Some of the compounds of this invention have asymmetric carbon atoms and therefore are enantiomers or diastereomers. Diasteromeric mixtures can be separated into their individual diastereomers on the basis of their physical chemical differences by methods known per se., for example, by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diasteromeric mixture by reaction with an appropriate optically active compound (e.g., alcohol), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. All such isomers, including diastereomers, enantiomers and mixtures thereof are considered as part of this invention. Also, some of the compounds of this invention are atropisomers (e.g., substituted biaryls) and are considered as part of this invention.
Those skilled in the art will recognize that the compounds of Formula I can exist in several tautomeric forms. All such tautomeric forms are considered as part of this invention. For example, all of the tautomeric forms of the carbonylguanidine moiety of the compounds of Formula I are included in this invention. Also, for example all enol-keto forms of the compounds of Formula I are included in this invention.
Some of the compounds of this invention are acidic and they form a salt with a pharmaceutically acceptable cation. All of the compounds of this invention are basic and they form a salt with a pharmaceutically acceptable anion. All such salts, including di-salts are within the scope of this invention and they can be prepared by conventional methods. For example, they can be prepared simply by contacting the acidic and basic entities, in either an aqueous, non-aqueous or partially aqueous medium. The salts are recovered either by filtration, by precipitation with a non-solvent followed by filtration, by evaporation of the solvent, or, in the case of aqueous solutions, by lyophilization, as appropriate.
In addition, when the compounds of this invention form metabolites, hydrates or solvates they are also within the scope of the invention.
Other cardiovascular agents known to those skilled in the art for example xcex2-blockers (e.g., acebutolol, atenolol, bopindolol, labetolol, mepindolol, nadolol, oxprenol, pindolol, propranolol, sotalol), calcium channel blockers (e.g., amlodipine, nifedipine, nisoldipine, nitrendipine, verapamil), potassium channel openers, adenosine, adenosine agoinists, ACE inhibitors (e.g., captopril, enalapril), nitrates (e.g., isosorbide dinitrate, isosorbide 5-mononitrate, glyceryl trinitrate), diuretics (e.g., hydrochlorothiazide, indapamide, piretanide, xipamide), glycosides (e.g., digoxin, metildigoxin), thrombolytics (e.g. tPA), platelet inhibitors (e.g., reopro), aspirin, dipyridamol, potassium chloride, clonidine, prazosin, aldose reductase inhibitors (e.g., zopolrestat) and adenosine A3 receptor agonists may be used in conjunction with the compounds of this invention.
In combination therapy treatment, both the compounds of this invention and the other drug therapies are administered to mammals (e.g., humans, male or female) by conventional methods.
Any aldose reductase inhibitor may be used as the second compound (active agent) of this invention for combination therapies. The term aldose reductase inhibitor refers to compounds which inhibit the bioconversion of glucose to sorbitol catalyzed by the enzyme aldose reductase. Such inhibition is readily determined by those skilled in the art according to standard assays (J. Malone, Diabetes, 29:861-864, 1980. xe2x80x9cRed Cell Sorbitol, an Indicator of Diabetic Controlxe2x80x9d). A variety of aldose reductase inhibitors are described and referenced below, however, other aldose reductase inhibitors will be known to those skilled in the art. The disclosures of U.S. patents listed below are hereby incorporated by reference. Also, common chemical USAN names or other designation are in parentheses where applicable, together with reference to appropriate patent literature disclosing the compound.
The activity of an aldose reductase inhibitor in a tissue can be determined by testing the amount of aldose reductase inhibitor that is required to lower tissue sorbitol (i.e., by inhibiting the further production of sorbitol consequent to blocking aldose reductase) or lower tissue fructose (by inhibiting the production of sorbitol consequent to blocking aldose reductase and consequently the production of fructose). While not wishing to be bound by any particular theory or mechanism, it is believed that an aldose reductase inhibitor, by inhibiting aldose reductase, prevents or reduces ischemic damage as described hereinafter.
Accordingly, examples of aldose reductase inhibitors useful in the compositions and methods of this invention include:
1. 3-(4-bromo-2-fluorobenzyl)-3,4-dihydro-4-oxo-1-phthalazineacetic acid (ponalrestat, U.S. Pat. No. 4,251,528);
2. N[[(5-trifluoromethyl)-6-methoxy-1-naphthalenyl]thioxomethyl}-N-methylglycine (tolrestat, U.S. Pat. No. 4,600,724);
3. 5-[(Z,E)-xcex2-methylcinnamylidene]-4-oxo-2-thioxo-3-thiazolideneacetic acid (epalrestat, U.S. Pat. Nos. 4,464,382, 4,791,126, 4,831,045);
4. 3-(4-bromo-2-fluorobenzyl)-7-chloro-3,4-dihydro-2,4-dioxo-1(2H)-quinazolineacetic acid (zenarestat, U.S. Pat. Nos. 4,734,419, and 4,883,800);
5. 2R,4R-6,7-dichloro-4-hydroxy-2-methylchroman-4-acetic acid (U.S. Pat. No. 4,883,410);
6. 2R,4R-6,7-dichloro-6-fluoro-4-hydroxy-2-methylchroman-4-acetic acid (U.S. Pat. No. 4,883,410);
7. 3,4-dihydro-2,8-diisopropyl-3-oxo-2H-1,4-benzoxazine-4-acetic acid (U.S. Pat. No. 4,771,050);
8. 3,4-dihydro-3-oxo-4-[(4,5,7-trifluoro-2-benzothiazolyl)methyl]-2H-1,4-benzothiazine-2-acetic acid (SPR-210, U.S. Pat. No. 5,252,572);
9. N-[3,5-dimethyl-4-[(nitromethyl)sulfonyl]phenyl]-2-methyl-benzeneacetamide (ZD5522, U.S. Pat. Nos. 5,270,342 and 5,430,060);
10. (S)-6-fluorospiro[chroman-4,4xe2x80x2-imidazolidine]-2,5xe2x80x2-dione (sorbinil, U.S. Pat. No. 4,130,714);
11. d-2-methyl-6-fluoro-spiro(chroman4xe2x80x2,4xe2x80x2-imidazolidine)-2xe2x80x2,5xe2x80x2-dione (U.S. Pat. No. 4,540,704);
12. 2-fluoro-spiro(9H-fluorene-9,4xe2x80x2imidazolidine)2xe2x80x2,5xe2x80x2-dione (U.S. Pat. No. 4,438,272);
13. 2,7-di-fluoro-spiro(9H-fluorene-9,4xe2x80x2imidazolidine)2xe2x80x2,5xe2x80x2-dione (U.S. Pat. Nos. 4,436,745, 4,438,272);
14. 2,7-di-fluoro-5-methoxy-spiro(9H-fluorene-9,4xe2x80x2imidazolidine)2xe2x80x2,5xe2x80x2-dione (U.S. Pat. Nos. 4,436,745, 4,438,272);
15. 7-fluoro-spiro(5H-indenol[1,2-b]pyridine-5,3xe2x80x2-pyrrolidine)2,5xe2x80x2-dione (U.S. Pat. Nos. 4,436,745, 4,438,272);
16. d-cis-6xe2x80x2-chloro-2xe2x80x2,3xe2x80x2-dihydro-2xe2x80x2-methyl-spiro-(imidazolidine-4,4xe2x80x2-4xe2x80x2-H-pyrano (2,3-b)pyridine)-2,5-dione (U.S. Pat. No. 4,980,357);
17. spiro[imidazolidine-4,5xe2x80x2(6H)-quinoline]2,5-dione-3xe2x80x2-chloro-7xe2x80x2,8xe2x80x2-dihydro-7xe2x80x2-methyl-(5xe2x80x2-cis) (U.S. Pat. No. 5,066,659);
18. (2S,4S)-6-fluoro-2xe2x80x2,5xe2x80x2-dioxospiro(chroman-4,4xe2x80x2-imidazolidine)-2-carboxamide (U.S. Pat. No. 5,447,946); and
19. 2-[(4-bromo-2-fluorophenyl)methyl]-6-fluorospiro[isoquinoline-4(1H),3xe2x80x2-pyrrolidine]-1,2xe2x80x2,3,5xe2x80x2(2H)-tetrone (ARI-509, U.S. Pat. No. 5,037,831).
Other aldose reductase inhibitors include compounds having formula IB 
or a pharmaceutically acceptable salt thereof, wherein
Z is O or S;
R1 is hydroxy or a group capable of being removed in vivo to produce a compound of formula IB wherein R1 is OH; and
X and Y are the same or different and are selected from hydrogen, trifluornomethyl, fluoro, and chloro.
A preferred subgroup within the above group of aldose reductase inhibitors includes numbered compounds 1, 2, 3, 4, 5, 6, 9, 10, and 17, and the following compounds of Formula IB:
20. 3,4-dihydro-3-(5-fluorobenzothiazol-2-ylmethyl)-4oxophthalazin-1-yl-acetic acid [R1=hydroxy; X=F; Y=H];
21. 3-(5,7-difluorobenzothiazol-2-ylmethyl)-3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1=hydroxy; X=Y=F];
22. 3-(5-chlorobenzothiazol-2-ylmethyl)-3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1=hydroxy; X=Cl; Y=H];
23. 3-(5,7-dichlorobenzothiazol-2-ylmethyl)-3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1=hydroxy; X=Y=Cl];
24. 3,4-dihydro-4-oxo-3-(5-trifluoromethylbenzoxazol-2-ylmethyl)phthalazin-1-ylacetic acid [R1=hydroxy; X=CF3; Y=H];
25. 3,4-dihydro-3-(5-fluorobenzoxazol-2-ylmethyl)-4-oxophthalazin-1-yl-acetic acid [R1=hydroxy; X=F; Y=H];
26. 3-(5,7-difluorobenzoxazol-2-ylmethyl)-3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1=hydroxy; X=Y=F];
27. 3-(5-chlorobenzoxazol-2-ylmethyl)-3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1=hydroxy; X=Cl; Y=H];
28. 3-(5,7-dichlorobenzoxazol-2-ylmethyl)-3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1=hydroxy; X=Y=Cl]; and
29. zopolrestat; 1-phthalazineacetic acid, 3,4-dihydro-4-oxo-3-[[5-(trifluoromethyl)-2-benzothiazolyl]methyl]-[R1=hydroxy; X=trifluoromethyl; Y=H].
In compounds 20-23, and 29 Z is S. In compounds 24-28, Z is O.
Of the above subgroup, compounds 20-29 are more preferred with 29 especially preferred.
An especially preferred aldose reductase inhibitor is 1-phthalazineacetic acid, 3,4-dihydro-4-oxo-3-[[5-trifluoromethyl)-2-benzothiazolyl]methyl]-.
The aldose reductase inhibitor compounds of this invention are readily available or can be easily synthesized by those skilled in the art using conventional methods of organic synthesis, particularly in view of the pertinent patent specification descriptions.
An amount of the aldose reductase inhibitor of this invention that is effective for the activities of this invention may be used. Typically, an effective dosage for the aldose reductase inhibitors of this invention is in the range of about 0.1 mg/kg/day to 100 mg/kg/day in single or divided doses, preferably 0.1 mg/kg/day to 20 mg/kg/day in single or divided doses.
Any glycogen phosphorylase inhibitor may be used as the second compound of this invention. The term glycogen phosphorylase inhibitor refers to any substance or agent or any combination of substances and/or agents which reduces, retards, or eliminates the enzymatic action of glycogen phosphorylase. The currently known enzymatic action of glycogen phosphorylase is the degradation of glycogen by catalysis of the reversible reaction of a glycogen macronmolecule and inorganic phosphate to glucose-1-phosphate and a glycogen macromolecule which is one glucosyl residue shorter than the original glycogen macromolecule (forward direction of glycogenolysis). Such actions are readily determined by those skilled in the art according to standard assays (e.g., as described hereinafter). A variety of these compounds are included in the following published international patent applications: PCT application publication WO 96/39384 and WO96/39385.However, other glycogen phosphorylase inhibitors will be known to those skilled in the art.
Preferred glycogen phosphorylase inhibitors include compounds having the Formula IC 
and the pharmaceutically acceptable salts and prodrugs thereof
wherein
the dotted line (---) is an optional bond;
A is xe2x80x94C(H)xe2x95x90, xe2x80x94C((C1-C4)alkyl)xe2x95x90 or xe2x80x94C(halo)xe2x95x90 when the dotted line (---) is a bond, or A is methylene or xe2x80x94CH((C1-C4)alkyl)xe2x80x94 when the dotted line (---) is not a bond;
R1, R10 or R11 are each independently H, halo, 4-, 6- or 7-nitro, cyano, (C1-C4)alkyl, (C1-C4)alkoxy, fluoromethyl, difluoromethyl or trifluoromethyl;
R2 is H;
R3 is H or (C1-C5)alkyl;
R4 is H, methyl, ethyl, n-propyl, hydroxy(C1-C3)alkyl, (C1-C3)alkoxy(C1-C3)alkyl, phenyl(C1-C4)alkyl, phenylhydroxy(C1-C4)alkyl, phenyl(C1-C4)alkoxy(C1-C4)alkyl, thien-2- or -3-yl(C1-C4)alkyl or fur-2- or -3-yl(C1-C4)alkyl wherein said R4 rings are mono-, di- or tri-substituted independently on carbon with H, halo, (C1-C4)alkyl, (C1-C4)alkoxy, trifluoromethyl, hydroxy, amino or cyano; or
R4 is pyrid-2-, -3- or -4-yl(C1-C4)alkyl, thiazol-2-, -4- or -5-yl(C1-C4)alkyl, imidazol -1-, -2-, -4- or -5-yl(C1-C4)alkyl, pyrrol-2- or -3-yl(C1-C4)alkyl, oxazol-2-, -4- or -5-yl-(C1-C4)alkyl, pyrazol-3-, 4- or -5-yl(C1-C4)alkyl, isoxazol-3-, -4- or -5-yl(C1-C4)alkyl, isothiazol-3-, -4- or -5-yl(C1-C4)alkyl, pyridazin-3- or -4-yl-(C1-C4)alkyl, pyrimidin-2-, -4-, -5- or -6-yl(C1-C4)alkyl, pyrazin-2- or -3-yl(C1-C4))alkyl or 1,3,5-triazin-2-yl(C1-C4)alkyl, wherein said preceding R4 heterocycles are optionally mono- or di-substituted independently with halo, trifluoromethyl, (C1-C4)alkyl, (C1-C4)alkoxy, amino or hydroxy and said mono- or di-substituents are bonded to carbon;
R5 is H, hydroxy, fluoro, (C1-C5)alkyl, (C1-C5)alkoxy, (C1-C6)alkanoyl, amino(C1-C4)alkoxy, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino(C1-C4)alkoxy, carboxy(C1-C4)alkoxy, (C1-C5)alkoxy-Carbonyl(C1-C4)alkoxy, benzyloxycarbonyl(C1-C4)alkoxy, or carbonyloxy wherein said carbonyloxy is carbon-carbon linked with phenyl, thiazolyl, imidazolyl, 1H-indolyl, furyl, pyrrolyl, oxazolyl, pyrazolyl, isoxazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl or 1,3,5-triazinyl and wherein said preceding R5 rings are optionally mono-substituted with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, amino or trifluoromethyl and said mono-substituents are bonded to carbon;
R7 is H, fluoro or (C1-C5)alkyl; or
R5 and R7 can be taken together to be oxo;
R6 is carboxy, (C1-C8)alkoxycarbonyl, C(O)NR8R9 or C(O)R12, wherein
R8 is H, (C1-C3)alkyl, hydroxy or (C1-C3)alkoxy; and
R9 is H, (C1-C8)alkyl, hydroxy, (C1-C8)alkoxy, methylene-perfluorinated(C1-C8)alkyl, phenyl, pyridyl, thienyl, furyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl, piperidinyl, morpholinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl or 1,3,5-triazinyl wherein said preceding R9 rings are carbon-nitrogen linked; or
R9 is mono-, di- or tri-substituted (C1-C5)alkyl, wherein said substituents are independently H, hydroxy, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C5)alkylamino; or
R9 is mono- or di-substituted (C1-C5)alkyl, wherein said substituents are independently phenyl, pyridyl, furyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl, pyridinyl, piperidinyl, morpholinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl or 1,3,5-triazinyl
wherein the nonaromatic nitrogen-Containing R9 rings are optionally mono-substituted on nitrogen with (C1-C6)alkyl, benzyl, benzoyl or (C1-C6)alkoxycarbonyl and wherein the R9 rings are optionally mono-substituted on carbon with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, amino, or mono-Nxe2x80x94 and di-N,N (C1-C5)alkylamino provided that no quaternized nitrogen is included and there are no nitrogen-oxygen, nitrogen-nitrogen or nitrogen-halo bonds;
R12 is piperazin-1-yl, 4-(C1-C4)alkylpiperazin-1-yl, 4-formylpiperazin-1-yl, morpholino, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxo-thiomorpholino, thiazolidin-3-yl, 1-oxo-thiazolidin-3-yl, 1,1-dioxo-thiazolidin-3-yl, 2-(C1-C6)alkoxycarbonylpyrrolidin-1-yl, oxazolidin-3-yl or 2(R)-hydroxymethylpyrrolidin-1-yl; or
R12 is 3- and/or 4-mono- or di-substituted oxazetidin-2-yl, 2-, 4-, and/or 5-mono- or di-substituted oxazolidin-3-yl, 2-, 4-, and/or 5- mono- or di-substituted thiazolidin-3-yl, 2-, 4-, and/or 5- mono- or di-substituted 1-oxothiazolidin-3-yl, 2-, 4-, and/or 5-mono- or di-substituted 1,1-dioxothiazolidin-3-yl, 3- and/or 4-, mono- or di-substituted pyrrolidin-1-yl, 3-, 4- and/or 5-, mono-, di- or tri-substituted piperidin-1-yl, 3-, 4-, and/or 5-mono-, di-, or tri-substituted piperazin-1-yl, 3-substituted azetidin-1-yl, 4- and/or 5-, mono- or di-substituted 1,2-oxazinan-2-yl, 3-and/or 4-mono- or di-substituted pyrazolidin-1-yl, 4- and/or 5-, mono- or di-substituted isoxazolidin-2-yl, 4- and/or 5-, mono- and/or di-substituted isothiazolidin-2-yl wherein said R12 substituents are independently H, halo, (C1-C5)-alkyl, hydroxy, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C5)alkylamino, formyl, oxo, hydroxyimino, (C1-C5)alkoxy, carboxy, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylcarbamoyl, (C1-C4)alkoxyimino, (C1-C4)alkoxymethoxy, (C1-C6)alkoxycarbonyl, carboxy(C1-C5)alkyl or hydroxy(C1-C5)alkyl;
with the proviso that if R4 is H, methyl, ethyl or n-propyl R5 is OH;
with the proviso that if R5 and R7 are H, then R4 is not H, methyl, ethyl, n-propyl, hydroxy(C1-C3)alkyl or (C1-C3)alkoxy(C1-C3)alkyl and R6 is C(O)NR8R9, C(O)R12 or (C1-C4)alkoxycarbonyl.
Preferred glycogen phosphorylase inhibitors include compounds having the Formula ID 
and the pharmaceutically acceptable salts and prodrugs thereof
wherein
the dotted line (---) is an optional bond;
A is xe2x80x94C(H)xe2x95x90, xe2x80x94C((C1-C4)alkyl)xe2x95x90, xe2x80x94C(halo)xe2x95x90 or xe2x80x94Nxe2x95x90, when the dotted line (---) is a bond, or A is methylene or xe2x80x94CH((C1-C4)alkyl)-, when the dotted line (---) is not a bond;
R1, R10 or R11 are each independently H, halo, cyano, 4-, 6-, or 7-nitro, (C1-C4)alkyl, (C1-C4)alkoxy, fluoromethyl, difluoromethyl or trifluoromethyl;
R2 is H;
R3 is H or (C1-C5)alkyl;
R4 is H, methyl, ethyl, n-propyl, hydroxy(C1-C3)alkyl, (C1-C3)alkoxy(C1-C3)alkyl, phenyl(C1-C4)alkyl, phenylhydroxy(C1-C4)alkyl, (phenyl)((C1-C4)-alkoxy)(C1-C4)alkyl, thien-2- or -3-yl(C1-C4)alkyl or fur-2- or -3-yl(C1-C4)alkyl wherein, said R4 rings are mono-, di- or tri-substituted independently on carbon with H, halo, (C1-C4)alkyl, (C1-C4)alkoxy, trifluoromethyl, hydroxy, amino, cyano or 4,5-dihydro-1H-imidazol-2-yl; or
R4 is pyrid-2-, -3- or 4-yl(C1-C4)alkyl, thiazol-2-, 4- or -5-yl(C1-C4)alkyl, imidazol-2-, -4- or -5-yl(C1-C4)alkyl, pyrrol-2- or -3-yl(C1-C4)alkyl, oxazol-2-, -4- or -5-yl(C1-C4)alkyl, pyrazol-3-, -4- or -5-yl(C1-C4)alkyl, isoxazol-3-, -4- or -5-yl(C1-C4)alkyl, isothiazol-3-, -4- or -5-yl(C1-C4)alkyl, pyridazin-3- or -4-yl(C1-C4)alkyl, pyrimidin-2-, -4-, -5- or -6-yl(C1-C4)alkyl, pyrazin-2- or -3-yl(C1-C4)alkyl, 1,3,5-triazin-2-yl(C1-C4)alkyl or indol-2-(C1-C4)alkyl, wherein said preceding R4 heterocycles are optionally mono- or di-substituted independently with halo, trifluoromethyl, (C1-C4)alkyl, (C1-C4)alkoxy, amino, hydroxy or cyano and said substituents are bonded to carbon; or
R4 is R15-carbonyloxymethyl, wherein said R15 is phenyl, thiazolyl, imidazolyl, 1H-indolyl, furyl, pyrrolyl, oxazolyl, pyrazolyl, isoxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or 1,3,5-triazinyl and wherein said preceding R15 rings are optionally mono- or di-substituted independently with halo, amino, hydroxy, (C1-C4)alkyl, (C1-C4)alkoxy or trifluoromethyl and said mono- or di-substituents are bonded to carbon;
R5 is H;
R6 is carboxy, (C1-C8)alkoxycarbonyl, benzyloxycarbonyl, C(O)NR8R9 or C(O)R12 wherein
R8 is H, (C1-C6)alkyl, cyclo(C3-C6)alkyl, cyclo(C3-C6)alkyl(C1-C5)alkyl, hydroxy or (C1-C8)alkoxy; and
R9 is H, cyclo(C3-C8)alkyl, cyclo(C3-C8)alkyl(C1-C5)alkyl, cyclo(C4-C7)alkenyl, cyclo(C3-C7)alkyl(C1-C5)alkoxy, cyclo(C3-C7)alkyloxy, hydroxy, methylene-perfluorinated(C1-C8)alkyl, phenyl, or a heterocycle wherein said heterocycle is pyridyl, furyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl, pyridinyl, piperidinyl, morpholinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, 1,3,5-triazinyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, thiochromanyl or tetrahydrobenzothiazolyl wherein said heterocycle rings are carbon-nitrogen linked; or
R9 is (C1-C6)alkyl or (C1-C8)alkoxy wherein said (C1-C6)alkyl or (C1-C8)alkdxy is optionally monosubstituted with cyclo(C4-C7)alken-1-yl, phenyl, thienyl, pyridyl, furyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl, piperidinyl, morpholinyl, thiomorpholinyl, 1-oxothiomorpholinyl, 1,1-dioxothiomorpholinyl, pyridazinyl, pyrmidinyl, pyrazinyl, piperazinyl, 1,3,5-triazinyl or indolyl and wherein said (C1-C6)alkyl or (C1-C8)alkoxy are optionally additionally independently mono- or di-substituted with halo, hydroxy, (C1-C5)alkoxy, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C5)alkylamino, cyano, carboxy, or (C1-C4)alkoxycarbonyl; and
wherein the R9 rings are optionally mono- or di-substituted independently on carbon with halo, (C1-C4)alkyl, (C1-C4)alkoxy, hydroxy, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino(C1-C4)alkyl, (C1-C4)alkoxy(C1-C4)alkyl, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino, cyano, carboxy, (C1-C5)alkoxycarbonyl, carbamoyl, formyl or trifluoromethyl and said R9 rings may optionally be additionally mono- or di-substituted independently with (C1-C5)alkyl or halo;
with the proviso that no quaternized nitrogen on any R9 heterocycle is included;
R12 is morpholino, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, thiazolidin-3-yl, 1-oxothiazolidin-3-yl, 1,1-dioxothiazolidin-3-yl, pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, piperazin-4-yl, azetidin-1-yl, 1,2-oxazinan-2-yl, pyrazolidin-1-yl, isoxazolidin-2-yl, isothiazolidin-2-yl, 1,2-oxazetidin-2-yl, oxazolidin-3-yl, 3,4-dihydroisoquinolin-2-yl, 1,3-dihydroisoindol-2-yl, 3,4-dihydro-2H-quinol-1-yl, 2,3-dihydro-benzo[1,4]oxazin-4-yl, 2,3-dihydro-benzo[1,4]-thiazine-4-yl, 3,4-dihydro-2H-quinoxalin-1-yl, 3,4-dihydro-benzo[c][1,2]oxazin-1-yl, 1,4-dihydro-benzo[d][1,2]oxazin-3-yl, 3,4-dihydro-benzo[e][1,2]-oxazin-2-yl, 3H-benzo[d]isoxazol-2-yl, 3H-benzo[c]isoxazol-1-yl or azepan-1-yl,
wherein said R12 rings are optionally mono-, di- or tri-substituted independently with halo, (C1-C5)alkyl, (C1-C5)alkoxy, hydroxy, amino, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C5)alkylamino, formyl, carboxy, carbamoyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C5)alkylcarbamoyl, (C1-C6)alkoxy(C1-C3)alkoxy, (C1-C5)alkoxycarbonyl, benzyloxycarbonyl, (C1-C5)alkoxycarbonyl(C1-C5)alkyl, (C1-C4)alkbxycarbonylamino, carboxy(C1-C5)alkyl, carbamoyl(C1-C5)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C5)alkylcarbamoyl(C1-C5)alkyl, hydroxy(C1-C5)alkyl, (C1-C4) alkoxy(C1-C4)alkyl, amino(C1-C4)alkyl, mono-Nxe2x80x94 or di-N,Nxe2x80x94(C1-C4)alkylamino(C1-C4)alkyl, oxo, hydroxyimino or (C1-C6)alkoxyimino and wherein no more than two substituents are selected from oxo, hydroxyimino or (C1-C6)alkoxyimino and oxo, hydroxyimino or (C1-C6)alkoxyimino are on nonaromatic carbon; and
wherein said R12 rings are optionally additionally mono- or di-substituted independently with (C1-C5)alkyl or halo;
with the proviso that when R6 is (C1-C5)alkoxycarbonyl or benzyloxycarbonyl then R1 is 5-halo, 5-(C1-C4)alkyl or 5-cyano and R4 is (phenyl)(hydroxy)(C1-C4)alkyl, (phenyl)((C1-C4)alkoxy)(C1-C4)alkyl, hydroxymethyl or Ar(C1-C2)alkyl, wherein Ar is thien-2- or -3-yl, fur-2- or -3-yl or phenyl wherein said Ar is optionally mono- or di-substituted independently with halo; with the provisos that when R4 is benzyl and R5 is methyl, R12 is not -4-hydroxy-piperidin-1-yl or when R4 is benzyl and R5 is methyl R6 is not C(O)N(CH3)2;
with the proviso that when R1 and R10 and R11 are H, R4 is not imidazol4-ylmethyl, 2-phenylethyl or 2-hydroxy-2-phenylethyl;
with the proviso that when both R8 and R9 are n-pentyl, R1 is 5-chloro, 5-bromo, 5-cyano, 5(C1-C5)alkyl, 5(C1-C5)alkoxy or trifluoromethyl;
with the proviso that when R12 is 3,4-dihydroisoquinol-2-yl, said 3,4-dihydroisoquinol-2-yl is not substituted with carboxy((C1-C4)alkyl;
with the proviso that when R8 is H and R9 is (C1-C6)alkyl, R9 is not substituted with carboxy or (C1-C4)alkoxycarbonyl on the carbon which is attached to the nitrogen atom N of NHR9; and
with the proviso that when R6 is carboxy and R1, R10, R11 and R5 are all H, then R4 is not benzyl, H, (phenyl)(hydroxy)methyl, methyl, ethyl or n-propyl.
In general an effective dosage for the pharmacological combination compositions of this invention, for example the ischemic damage reducing activities of combinations containing the glycogen phosphorylase inhibitor compounds of this invention, is in the range of 0.005 to 50 mg/kg/day, preferably 0.01 to 25 mg/kg/day and most preferably 0.1 to 15 mg/kg/day.
The compounds of the present invention inhibit the sodium/proton (Na+/H+) exchange transport system and hence are useful as a therapeutic or prophylactic agent for diseases caused or aggravated by the acceleration of the sodium/proton (Na+/H+) exchange transport system, for example, cardiovascular diseases [e.g., arteriosclerosis, hypertension, arrhythmia (e.g. ischemic arrhythmia, arrhythmia due to myocardial infarction, myocardial stunning, myocardial dysfunction, arrhythmia after PTCA or after thrombolysis, etc.), angina pectoris, cardiac hypertrophy, myocardial infarction, heart failure (e.g. congestive heart failure, acute heart failure, cardiac hypertrophy, etc.), restenosis after PTCA, PTCI, shock (e.g. hemorrhagic shock, endotoxin shock, etc.)], renal diseases (e.g. diabetes mellitus, diabetic nephropathy, ischemic acute renal failure, etc.) organ disorders associated with ischemia or ischemic reperfusion [(e.g. heart muscle ischemic reperfusion associated disorders, acute renal failure, or disorders induced by surgical treatment such as coronary artery bypass grafting (CABG) surgeries, vascular surgeries, organ transplantation, non-Cardiac surgeries or percutaneous transluminal coronary angioplasty (PTCA)], cerebrovascular diseases (e.g. ischemic stroke, hemorrhagic stroke, etc.), cerebro ischemic disorders (e.g. disorders associated with cerebral infarction, disorders caused after cerebral apoplexy as sequelae, or cerebral edema. The compounds of this invention can also be used as an agent for myocardial protection during coronary artery bypass grafting (CABG) surgeries, vascular surgeries, percutaneous transluminal coronary angioplasty (PTCA), PTCI, organ transplantation, or non-Cardiac surgeries.
Preferably, the compounds of this invention can be used as agents for myocardial protection before, during, or after coronary artery bypass grafting (CABG) surgeries, vascular surgeries, percutaneous transluminal coronary angioplasty (PTCA), organ transplantation, or nonrdiac surgeries.
Preferably, the compounds of this invention can be used as agents for myocardial protection in patients presenting with ongoing cardiac (acute coronary syndromes, e.g. myocardial infarction or unstable angina) or cerebral ischemic events (e.g. stroke).
Preferably, the compounds of this invention can be used as agents for chronic myocardial protection in patients with diagnosed coronary heart disease (e.g. previous myocardial infarction or unstable angina) or patients who are at high risk for myocardial infarction (age greater than 65 and two or more risk factors for coronary heart disease).
In addition to this, the compounds of this invention are notable for their strong inhibitory effect on the proliferation of cells, for example the proliferation of fibroblast cells and the proliferation of the smooth muscle cells of the blood vessels. For this reason, the compounds of this invention are valuable therapeutic agents for use in diseases in which cell proliferation represents a primary or secondary cause and may, therefore, be used as antiatherosclerotic agents, and as agents against diabetic late complications, cancerous diseases, fibrotic diseases such as pulmonary fibrosis, hepatic fibrosis or renal fibrosis, glomerular nephrosclerosis, organ hypertrophies or hyperplasias, in particular hyperplasia or hypertrophy of the prostate, pulmonary fibrosis, diabetic complications or recurrent stricture after PTCA, or diseases caused by endothelial cell injury.
The utility of the compounds of the present invention as medical agents in the treatment of diseases, such as are detailed herein in mammals (e.g. humans) for example, myocardial protection during surgery or mycardial protection in patients presenting with ongoing cardiac or cerebral ischemic events or chronic cardioprotection in patients with diagnosed coronary heart disease, or at risk for coronary heart disease, cardiac dysfunction or myocardial stunning is demonstrated by the activity of the compounds of this invention in conventional preclinical cardioprotection assays [see the in vivo assay in Klein, H. et al., Circulation 92:912-917 (1995); the isolated heart assay in Scholz, W. et al., Cardiovascular Research 29:260-268 (1995); the antiarrhythmic assay in Yasutake M. et al., Am. J. Physiol., 36:H2430-H2440 (1994); the NMR assay in Kolke et al., J. Thorac. Cardiovasc. Surg. 112: 765-775 (1996)] and the additional in vitro and in vivo assays described below. Such assays also provide a means whereby the activities of the compounds of this invention can be compared with the activities of other known compounds. The results of these comparisons are useful for determining dosage levels in mammals, including humans, for the treatment of such diseases.
Methodologies for measurement of human NHE-1 activity and inhibitor potency are based on those published by Watson et al., Am. J. Physiol., 24:G229-G238, 1991), where NHE-mediated recovery of intracellular pH is measured following intracellular acidification. Thus, fibroblasts stably expressing human NHE-1 (Counillon, L. et al., Mol. Pharmacol., 44:1041-1045 (1993) are plated onto collagen coated 96 well plates (50,000/well) and grown to confluence in growth media (DMEM high glucose, 10% fetal bovine serum, 50 u/ml penicillin and streptomycin). Confluent plates are incubated for 30 min at 37xc2x0 C. with the pH sensitive fluorescent probe BCECF (5 xcexcM; Molecular Probes, Eugene, Oreg.). BCECF loaded cells are incubated for 30 min at 37xc2x0 C. in acid loading media (70 mM choline chloride, 50 mM NHCl4, 5 mM KCl, 1 mM MgCl2, 1.8 mM CaCl2, 5 mM glucose, 10 mM HEPES, pH 7.5), and then placed in a Fluorescent Imaging Plate Reader (Molecular Devices, CA). BCECF fluorescence is monitored using excitation and emission wavelengths of 485 nM and 525 nM, respectively. Intracellular acidification is initiated via rapid replacement of acid loading media with recovery media (120 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1.8 mM CaCl2, 5 mM glucose, 10 mM HEPES, pH 7.5)xc2x1test compound, and NHE-mediated recovery of intracellular pH is monitored as the subsequent time-dependent increase BCECF fluorescence. The potency of human NHE-1 inhibitors is calculated as the concentration that reduces recovery of intracellular pH by 50% (IC50). Under these conditions reference NHE inhibitors amiloride and HOE-642 had IC50 values for human NHE-1 of 50 xcexcM and 0.5 xcexcM, respectively.
As background information, it is noted that brief periods of myocardial ischemia followed by coronary artery reperfusion protects the heart from subsequent severe myocardial ischemia (Murry et al., Circulation 74:1124-1136, 1986).
The therapeutic effects of the compounds of this invention in preventing heart tissue damage resulting from an ischemic insult can be demonstrated in vitro along lines presented in Liu et al. (Cardiovasc. Res., 28:1057-1061, 1994), as described specifically herein. Cardioprotection, as indicated by a reduction in infarcted myocardium, can be induced pharmacologically using adenosine receptor agonists in isolated, retrogradely perfused rabbit hearts as an in vitro model of myocardial ischemic preconditioning (Liu et al., Cardiovasc. Res., 28:1057-1061, 1994). The in vitro test described below demonstrates that a test compound (i.e., a compound as claimed herein) can also pharmacologically induce cardioprotection, i.e., reduced myocardial infarct size, when administered to a rabbit isolated heart. The effects of the test compound are compared to ischemic preconditioning and the A1/A3 adenosine agonist, APNEA (N6-[2-(4-aminophenyl)ethyl]adenosine), that has been shown to pharmacologically induce cardioprotection in the rabbit isolated heart (Liu et al., Cardiovasc. Res., 28:1057-1061, 1994). The exact methodology is described below.
The protocol used for these experiments closely follows that described by Liu et al., Cardiovasc. Res., 28:1057-1061, 1994. Male New Zealand White rabbits (3-4 kg) are anesthetized with sodium pentobarbital (30 mg/kg, i.v.). After deep anesthesia is achieved (determined by the absence of an ocular blink reflex) the animal is intubated and ventilated with 100% O2 using a positive pressure ventilator. A left thoracotomy is performed, the heart exposed, and a snare (2-0 silk) is placed loosely around a prominent branch of the left coronary artery, approximately ⅔ of the distance towards the apex of the heart. The heart is removed from the chest and rapidly ( less than 30 sec) mounted on a Langendorff apparatus. The heart is retrogradely perfused in a non-recirculating manner with a modified Krebs solution (NaCl 118.5 mM, KCl 4.7 mM, Mg SO4 1.2 mM, KH2PO4 1.2 mM, NaHCO3 24.8 mM, CaCl2 2.5 mM, and glucose 10 mM), at a constant pressure of 80 mmHg and a temperature of 37xc2x0 C. Perfusate pH is maintained at 7.4-7.5 by bubbling with 95% O2/5% CO2. Heart temperature is tightly controlled by using heated reservoirs for the physiological solution and water jacketing around both the perfusion tubing and the isolated heart. Heart rate and left ventricular pressures are determined via a latex balloon which is inserted in the left ventricle and connected by stainless steel tubing to a pressure transducer. The intraventricular balloon is inflated to provide a systolic pressure of 80-100 mmHg, and a diastolic pressure xe2x89xa610 mmHg. Total coronary flow is also continuously monitored using an in-line flow probe and normalized for heart weight.
The heart is allowed to equilibrate for 30 min, over which time the heart must show stable left ventricular pressures within the parameters outlined above. If the heart rate falls below 180 bpm at any time prior to the 30 min period of regional ischemia, the heart is paced at about 200 bpm for the remainder of the experiment. Ischemic preconditioning is induced by total cessation of cardiac perfusion (global ischemia) for 5 min, followed by reperfusion for 10 min. The regional ischemia is provided by tightening the snare around the coronary artery branch. Following the 30 min regional ischemia, the snare is released and the heart reperfused for an additional 120 min.
Pharmacological cardioprotection is induced by infusing the test compound at predetermined concentrations, starting 30 min prior to the 30 min regional ischemia, and continuing until the end of the 120 min reperfusion period. Hearts which receive test compounds do not undergo the period of ischemic preconditioning. The reference compound, APNEA (500 nM) is perfused through hearts (which do not receive the test compound) for a 5 min period which ends 10 min before the 30 min regional ischemia.
At the end of the 120 min reperfusion period, the coronary artery snare is tightened, and a 0.5% suspension of fluorescent zinc cadmium sulfate particles (1-10 xcexcM) Duke Scientific Corp. (Palo Alto, Calif.) is perfused through the heart; this stains all of the myocardium, except that area-at-risk for infarct development (area-at-risk). The heart is removed from the Langendorff apparatus, blotted dry, wrapped in aluminum foil and stored overnight at xe2x88x9220xc2x0 C. The next day, the heart is sliced into 2 mm transverse sections from the apex to the top of the ventricles. The slices are stained with 1% triphenyl tetrazolium chloride (TTC) in phosphate-buffered saline for 20 min at 37xc2x0 C. Since TTC reacts with living tissue (containing NAD-dependent dehydrogenase), this stain differentiates between living (red stained) tissue, and dead tissue (unstained infarcted tissue). The infarcted area (no stain) and the area-at-risk (no fluorescent particles) are calculated for each slice of left ventricle using a precalibrated image analyzer. To normalize the ischemic injury for differences in the area-at-risk between hearts, the data is expressed as the ratio of infarct area vs. area-at-risk (%IA/AAR). All data are expressed as meanxc2x1SE and compared statistically using a Mann-Whitney non-parametric test with a Bonferroni correction for multiple comparisons. Significance is considered as p less than 0.05.
The results from the above in vitro test demonstrate that compounds of this invention induce significant cardioprotection relative to the control group.
The therapeutic effects of the compounds of this invention in preventing heart tissue damage otherwise resulting from an ischemic insult can also be demonstrated in vivo along lines presented in Liu et al. (Circulation, Vol. 84:350-356, 1991) as described specifically herein. The in vivo assay tests the cardioprotection of the test compound relative to the control group which receives saline vehicle. Cardioprotection, as indicated by a reduction in infarcted myocardium, can be induced pharmacologically using intravenously administered adenosine receptor agonists in intact, anesthetized rabbits studied as an in situ model of myocardial ischemic preconditioning (Liu et al., Circulation 84:350-356, 1991). The in vivo assay tests whether compounds can pharmacologically induce cardioprotection, i.e., reduced myocardial infarct size, when parenterally administered to intact, anesthetized rabbits. The effects of the compounds of this invention can be compared to ischemic preconditioning using the A1 adenosine agonist, N6-1-(phenyl-2R-isopropyl) adenosine (PIA) that has been shown to pharmacologically induce cardioprotection in intact anesthetized rabbits studied in situ (Liu et al., Circulation 84:350-356, 1991). The methodology is described below.
Surgery: New Zealand White male rabbits (3-4 kg) are anesthetized with sodium pentobarbital (30 mg/kg, i.v.). A tracheotomy is performed via a ventral midline cervical incision and the rabbits are ventilated with 100% oxygen using a positive pressure ventilator. Catheters are placed in the left jugular vein for drug administration and in the left carotid artery for blood pressure measurements. The hearts are then exposed through a left thoracotomy and a snare (00 silk) placed around a prominent branch of the left coronary artery. Ischemia is induced by pulling the snare tight and clamping it in place. Releasing the snare allows the affected area to reperfuse. Myocardial ischemia is evidenced by regional cyanosis; reperfusion is evidenced by reactive hyperemia.
Protocol: Once arterial pressure and heart rate have been stable for at least 30 minutes the test is started. Ischemic preconditioning is induced by occluding the coronary artery for 5 min followed by a 10 min reperfusion. Pharmacological preconditioning is induced by infusing test compound over, for example 5 minutes and allowing 10 minutes before further intervention or by infusing the adenosine agonist, PIA (0.25 mg/kg). Following ischemic preconditioning, pharmacological preconditioning or no conditioning (unconditioned, vehicle control) the artery is occluded for 30 minutes and then reperfused for two hours to induce myocardial infarction. The test compound and PIA are dissolved in saline or other suitable vehicle and delivered at 1 to 5 mg/kg, respectively.
Staining (Liu et al., Circulation 84:350-356, 1991): At the end of the 2 hour reperfusion period, the hearts are quickly removed, hung on a Langendorff apparatus, and flushed for 1 minute with normal saline heated to body temperature (38xc2x0 C.). The silk suture used as the snare is then tied tightly to reocclude the artery and a 0.5% suspension of fluorescent zinc cadmium sulphate particles (1-10 xcexcm) Duke Scientific Corp. (Palo Alto, Calif.) is infused with the perfusate to stain all of the myocardium except the area at risk (nonfluorescent ventricle). The hearts are then quickly frozen and stored overnight at xe2x88x9220xc2x0 C. On the following day, the hearts are cut into 2 mm slices and stained with 1% triphenyl tetrazolium chloride (TTC). Since TTC reacts with living tissue, this stain differentiates between living (red stained) tissue, and dead tissue (unstained infarcted tissue). The infarcted area (no stain) and the area at risk (no fluorescent particles) are calculated for each slice of left ventricle using a pre-calibrated image analyzer. To normalize the ischemic injury for differences in the area at risk between hearts, the data is expressed as the ratio of infarct area vs. area at risk (%IA/AAR). All data are expressed as Meanxc2x1SEM and compared statistically using single factor ANOVA or Mann Whitney non parametric test. Significance is considered as p less than 0.05.
The compounds of this invention can be tested for their utility in reducing or preventing ischemic injury in non-cardiac tissues, for example, the brain, or the liver, utilizing procedures reported in the scientific literature. The compounds of this invention in such tests can be administered by the preferred route and vehicle of administration and at the preferred time of administration either prior to the ischemic episode, during the ischemic episode, following the ischemic episode (reperfusion period) or during any of the below-mentioned experimental stages.
The benefit of the invention to reduce ischemic brain damage can be demonstrated, for example, in mammals using the method of Park, et al (Ann. Neurol. 1988;24:543-551). According to the procedure of Park, et al., adult male Sprague Dawley rats are anesthetized initially with 2% halothane, and thereafter by mechanical ventilation with a nitrous oxide-oxygen mixture (70%:30%) containing 0.5-1% halothane. A tracheostomy is then performed. The stroke volume of the ventilator is adjusted to maintain arterial carbon dioxide tension at approximately 35 mm Hg and adequate arterial oxygenation (PaO2 greater than 90 mm Hg). Body temperature can be monitored by a rectal thermometer, and the animals can be maintained normothermic, if necessary, by external heating. The animals next undergo subtemporal craniectomy to expose the main trunk of the left middle cerebral artery (MCA) under an operating microscope, and the exposed artery is occluded with microbipolar coagulation to generate large ischemic lesions in the cerebral cortex and basal ganglia. After three hours of MCA occlusion, the rats are deeply anesthetized with 2% halothane and a thoracotomy is performed to infuse heparinized saline into the left ventricle. The effluent is collected via an incision of the right atrium. The saline washout is followed by approximately 200 ml of a 40% formaldehyde, glacial acetic acid and absolute methanol solution (FAM; 1:1:8, v/v/v), then the animals are decapitated and the head is stored in fixative for 24 hours. The brain is then removed, dissected, embedded in paraffin wax, and sectioned (approximately 100 sections 0.2 mm per brain). The sections are then stained with hematoxylin-eosin or with a combination of cresyl violet and Luxol fast blue, and examined by light microscopy to identify and quantitate the ischemic damage using a precalibrated image analyzer. The ischemic volumes and areas are expressed in absolute units (mm3 and mm2) and as a percentage of the total region examined. The effect of the compositions and methods of this invention to reduce ischemic brain damage induced by MCA occlusion is noted based on a reduction in the area or volume of relative or absolute ischemic damage in the brain sections from the rats in the treatment group compared to brain sections from rats in a placebo-treated control group.
Other methods which could alternatively be utilized to demonstrate the benefit of the invention to reduce ischemic brain damage include those described by Nakayama, et al. in Neurology 1988,38:1667-1673; Memezawa, et al. in Stroke 1992,23:552-559; Folbergrova, et al. in Proc. Natl. Acad. Sci 1995,92:5057-5059; and Gotti, et al. in Brain Res. 1990,522:290-307.
The benefit of the compounds, compositions and methods of this invention to reduce ischemic liver damage can be demonstrated, for example, in mammals using the method of Yokoyama, et al. (Am. J. Physiol. 1990;258:G564-G570). According to the procedure of Yokoyama, et al., fasted adult male Sprague Dawley rats are anesthetized with sodium pentobarbital (40 mg/kg i.p.), then the animals are tracheotomized and mechanically ventilated with room air. The liver is extirpated and placed in an environmental chamber maintained at constant temperature (37xc2x0 C.), then perfused through the portal vein at a constant pressure of 15 cm H2O with a modified, hemoglobin-free Krebs-Henseleit buffer (in mM: 118 NaCl, 4.7 KCl, 27 NaHCO3, 2.5 CaCl2, 1.2 MgSO4, 1.2 KH2PO4, 0.05 EDTA, and 11 mM glucose, plus 300 U heparin). The pH of the perfusate is maintained at 7.4 by gassing the buffer with 95% O2xe2x80x945% CO2. Each liver is perfused at a flow rate of 20 ml/min in a single-pass manner for a 30 min washout and equilibration period (preischemic period), followed by a 2 hour period of global ischemia, and then a 2 hour period of reperfusion under conditions identical to the preischemic period. Aliquots (20 ml) of the perfusate are collected during the preischemic period, immediately after the occlusive ischemic period, and every 30 min of the 2 hour reperfusion period. The perfusate samples are assayed for the appearance of hepatocellular enzymes, for example, aspartate amino-transferase (AST), alanine amino4transferase (ALT), and lactate dehydrogenase (LDH), which are taken to quantitatively reflect the degree of ischemic liver tissue damage during the procedure. AST, ALT, and LDH activities in the perfusate can be determined by several methods, for example, by the reflectometry method using an automatic Kodak Ektachem 500 analyzer reported by Nakano, et al. (Hepatology 1995;22:539-545). The effect of the compounds, compositions and methods of this invention in reducing ischemic liver damage induced by occlusion is noted based on a reduction in the release of hepatocellular enzymes immediately following the occlusive period and/or during the postischemic-reperfusion period in the perfused livers from the rats in the treatment group compared to perfused livers from rats in a placebo-treated control group.
Other methods and parameters which could alternatively be utilized to demonstrate the benefit of the compositions and methods of this invention in reducing ischemic liver damage include those described by Nakano, et al. (Hepatology 1995;22:539-545).
Male Sprague-Dawley rats are rendered diabetic by injection of streptozocin at 55 mg/kg, i.v., in pH 4.5 citrate buffer. They are fed ad libitum in controlled conditions of housing, temperature and lighting. After five weeks of diabetes, the rats are anesthetized with an overdose of pentobarbital, and tissues are rapidly removed and analyzed for sorbitol and fructose.
Sorbitol levels are analyzed according to the method of Donald M. Eades et al., xe2x80x9cRapid Analysis of Sorbitol, Galactitol, Mannitol and Myoinositol Mixtures From Biological Sourcesxe2x80x9d, Journal of Chromatography, 490, 1-8, (1989).
Fructose in rat tissues is enzymatically measured using a modification of the method of Ameyama (Methods in Enzymology, 89:20-29, 1982), in which ferricyanide was replaced by resazurin, a dye that is reduced to the highly fluorescent resorufin. The amount of resorufin fluorescence is stoichiometric with the amount of fructose oxidized by fructose dehydrogenase. The assay contains 0.1 ml neutralized 6% perchloric acid nerve extract in a final volume of 1.5 ml. Following incubation for 60 minutes at room temperature in a closed drawer, sample fluorescence is determined at excitation=560 nm, emission=580 nm with slits of 5 mm each in a Perkin-Elmer model 650-40 fluorescence spectrophotometer. Fructose concentrations are calculated by comparison with a series of known fructose standards.
The three different purified glycogen phosphorylase (GP) isoenzymes, wherein glycogen phosphorylase is in the activated xe2x80x9caxe2x80x9d state (referred to as glycogen phosphorylase a, or the abbreviation GPa), and referred to here as human liver glycogen phosphorylase a (HLGPa), human muscle glycogen phosphorylase a (HMGPa), and human brain glycogen phosphorylase a (HBGPa), can be obtained by the following procedures.
Expression and Fermentation
The HLGP and HMGP cDNAs are expressed from plasmid pKK233-2 (Pharmacia Biotech. Inc., Piscataway, N.J.) in E. coli strain XL-1 Blue (Stratagene Cloning Systems, LaJolla, Calif.). The strain is inoculated into LB medium (consisting of 10 g tryptone, 5 g yeast extract, 5 g NaCl, and 1 ml 1N NaOH per liter) plus 100 mg/L ampicillin, 100 mg/L pyridoxine and 600 mg/L MnCl2 and grown at 37xc2x00 C. to a cell density of OD550=1.0. At this point, the cells are induced with 1 mM isopropyl-1-thio-xcex2-D-galactoside (IPTG). Three hours after induction the cells are harvested by centrifugation and cell pellets are frozen at xe2x88x9270xc2x00 C. until needed for purification.
The HBGP cDNA can be expressed by several methodologies, for example, by the method described by Crerar, et al. (J. Biol. Chem. 270:13748-13756). The method described by Crerar, et al. (J. Biol. Chem. 270:13748-13756) for the expression of HBGP is as follows: the HBGP cDNA can be expressed from plasmid pTACTAC in E. Coli strain 25A6. The strain is inoculated into LB medium (consisting of 10 g tryptone, 5 g yeast extract, 5 g NaCl, and 1 ml 1N NaOH per liter) plus 50 mg/L ampicillin and grown overnight, then resuspended in fresh LB medium plus 50 mg/L ampicillin, and reinoculated into a 40xc3x97 volume of LB/amp media containing 250 xcexcM isopropyl-1-thio-xcex2-D-galactoside (IPTG), 0.5 mM pyridoxine and 3 mM MgCl2 and grown at 22xc2x0 C. for 48-50 hours. The cells can then be harvested by centrifugation and cell pellets are frozen at xe2x88x9270xc2x0 C. until needed for purification.
The HLGP cDNA is expressed from plasmid pBlueBac III (Invitrogen Corp., San Diego, Calif.) which is cotransfected with BaculoGold Linear Viral DNA (Pharmingen, San Diego, Calif.) into Sf9 cells. Recombinant virus is subsequently plaque-purified. For production of protein, Sf9 cells grown in serum-free medium are infected at a multiplicity of infection (moi) of 0.5 and at a cell density of 2xc3x97106 cells/ml. After growth for 72 hours at 27xc2x0 C., cells are centrifuged, and the cell pellets frozen at xe2x88x9270xc2x0 C. until needed for purification. Purification of Glycogen Phosphhorylase expressed in E. coli. 
The E. coli cells in pellets described above are resuspended in 25 mM xcex2-glycerophosphate (pH 7.0) with 0.2 mM DTT, 1 mM MgCl2, plus the following protease inhibitors:
lysed by pretreatment with 200 xcexcg/mL lysozyme and 3 xcexcg/mL DNAase followed by sonication in 250 mL batches for 5xc3x971.5 minutes on ice using a Branson Model 450 ultrasonic cell disrupter (Branson Sonic Power Co., Danbury Conn.). The E. coli cell lysates are then cleared by centrifugation at 35,000xc3x97g for one hour followed by filtration through 0.45 micron filters. GP in the soluble fraction of the lysates (estimated to be less than 1% of the total protein) is purified by monitoring the enzyme activity (as described in GPa Activity Assay section, below) from a series of chromatographic steps detailed below.
Immobilized Metal Affinity Chromatography (IMAC)
This step is based on the method of Luong et al (Luong et al. Joumal of Chromatography (1992) 584, 77-84.). 500 mL of the filtered soluble fraction of cell lysates (prepared from approximately 160-250 g of original cell pellet) are loaded onto a 130 mL column of IMAC Chelating-Sepharose (Pharmacia LKB Biotechnology, Piscataway, N.J.) which has been charged with 50 mM CuCl2 and 25 mM xcex2-glycerophosphate, 250 mM NaCl and 1 mM imidazole at pH 7 equilibration buffer. The column is washed with equilibration buffer until the A280 returns to baseline. The sample is then eluted from the column with the same buffer containing 100 mM imidazole to remove the bound GP and other bound proteins. Fractions containing the GP activity are pooled (approximately 600 mL), and ethylenediaminetetraacetic acid (EDTA), DL-dithiothreitol (DTT), phenylmethylsulfonyl fluoride (PMSF), leupeptin and pepstatin A are added to obtain 0.3 mM, 0.2 mM, 0.2 mM, 0.5 xcexcg/mL and 0.7 xcexcg/mL concentrations respectively. The pooled GP is desalted over a Sephadex G-25 column (Sigma Chemical Co., St. Louis, Mo.) equilibrated with 25 mM Tris-HCl (pH 7.3), 3 mM DTT buffer (Buffer A) to remove imidazole and is stored on ice until the second chromatographic step.
5xe2x80x2-AMP-Sepharose Chromatography
The desalted pooled GP sample (approximately 600 mL) is next mixed with 70 mL of 5xe2x80x2-AMP Sepharose (Pharmacia LKB Biotechnology, Piscataway, N.J.) which has been equilibrated with Buffer A (see above). The mixture is gently agitated for one hour at 22xc2x0 C. then packed into a column and washed with Buffer A until the A280 returns to baseline. GP and other proteins are eluted from the column with 25 mM Tris-HCl, 0.2 mM DTT and 10 mM adenosine 5xe2x80x2-monophosphate (AMP) at pH 7.3 (Buffer B). GP-containing fractions are pooled following identification by determining enzyme activity (described below) and visualizing the Mr approximately 97 kdal GP protein band by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by silver staining (2D-silver Stain II xe2x80x9cDaiichi Kitxe2x80x9d, Daiichi Pure Chemicals Co., LTD., Tokyo, Japan) and then pooled. The pooled GP is dialyzed into 25 mM xcex2-glycerophosphate, 0.2 mM DTT, 0.3 mM EDTA, 200 mM NaCl, pH 7.0 buffer (Buffer C) and stored on ice until use.
Prior to use of the GP enzyme, the enzyme is converted from the inactive form as expressed in E. coli strain XL-1 Blue (designated GPb) (Stragene Cloning Systems, La Jolla, Calif.), to the active form (designated GPa) by the procedure described in Section (A) Activation of GP below.
Purification of Glycogen Phosphorylase Expressed in Sf9 Cells
The Sf9 cells in pellets described above are resuspended in 25 mM xcex2-glycerophosphate (pH 7.0) with 0.2 mM DTT, 1 mM MgCl2, plus the following protease inhibitors:
lysed by pretreatment with 3 xcexcg/mL DNAase followed by sonication in batches for 3xc3x971 minutes on ice using a Branson Model 450 ultrasonic cell disrupter (Branson Sonic Power Co., Danbury Conn.). The Sf9 cell lysates are then cleared by centrifugation at 35,000xc3x97g for one hour followed by filtration through 0.45 micron filters. GP in the soluble fraction of the lysates (estimated to be 1.5% of the total protein) is purified by monitoring the enzyme activity (as described in GPa Activity Assay section, below) from a series of chromatographic steps detailed below.
Immobilized Metal Affinity Chromatography (IMAC)
Immobilized Metal Affinity Chromatography is performed as described in the section above. The pooled, desalted GP is then stored on ice until further processed.
Activation of GP
Before further chromatography, the fraction of inactive enzyme as expressed in Sf9 cells (designated GPb) is converted to the active form (designated GPa) by the following procedure described in Section (A) Activation of GP below.
Anion Exchange Chromatography
Following activation of the IMAC purified GPb to GPa by reaction with the immobilized phosphorylase kinase, the pooled GPa fractions are dialyzed against 25 mM Tris-HCl, pH 7.5, containing 0.5 mM DTT, 0.2 mM EDTA, 1.0 mM phenylmethylsulfonyl fluoride (PMSF), 1.0 xcexcg/mL leupeptin and 1.0 xcexcg/mL pepstatin A. The sample is then loaded onto a MonoQ Anion Exchange Chromatography column (Pharmacia Biotech. Inc., Piscataway, N.J.). The column is washed with equilibration buffer until the A280 returns to baseline. The sample is then eluted from the column with a linear gradient of 0-0.25 M NaCl to remove the bound GP and other bound proteins. GP-containing fractions elute between 0.1-0.2 M NaCl range, as detected by monitoring the eluant for peak protein absorbance at A280. The GP protein is then identified by visualizing the Mr approximately 97 kdal GP protein band by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by silver staining (2D-silver Stain II xe2x80x9cDaiichi Kitxe2x80x9d, Daiichi Pure Chemicals Co., LTD., Tokyo, Japan) and then pooled. The pooled GP is dialyzed into 25 mM N,N-bis[2-Hydroxyethyl]-2-aminoethanesulfonic acid, 1.0 mM DTT, 0.5 mM EDTA, 5 mM NaCl, pH 6.8 buffer and stored on ice until use.
Determination of GP Enzyme Activity
A) Activation of GP: Conversion of GPb to GPa
Prior to the determination of GP enzyme activity, the enzyme is converted from the inactive form as expressed in E. coli strain XL-1 Blue (designated GPb) (Stragene Cloning Systems, La Jolla, Calif.), to the active form (designated GPa) by phosphorylation of GP using phosphorylase kinase as follows. The fraction of inactive enzyme as expressed in Sf9 cells (designated GPb) is also converted to the active form (designated GPa) by the following procedure.
GP Reaction with Immobilized Phosphorylase Kinase
Phosphorylase kinase (Sigma Chemical Company, St. Louis, Mo.) is immobilized on Affi-Gel 10 (BioRad Corp., Melvile, N.Y.) as per the manufacturer""s instructions. In brief, the phosphorylase kinase enzyme (10 mg) is incubated with washed Affi-Gel beads (1 mL) in 2.5 mL of 100 mM HEPES and 80 mM CaCl2 at pH 7.4 for 4 hours at 4xc2x0 C. The Affi-Gel beads are then washed once with the same buffer prior to blocking with 50 mM HEPES and 1 M glycine methyl ester at pH 8.0 for one hour at room temperature. Blocking buffer is removed and replaced with 50 mM HEPES (pH 7.4), 1 mM xcex2-mercaptoethanol and 0.2% NaN3 for storage. Prior to use to convert GPb to GPa, the Affi-Gel immobilized phosphorylase kinase beads are equilibrated by washing in the buffer used to perform the kinase reaction, consisting of 25 mM xcex2-glycerophosphate, 0.3 mM DTT, and 0.3 mM EDTA at pH 7.8 (kinase assay buffer).
The partially purified, inactive GPb obtained from 5xe2x80x2-AMP-Sepharose chromatography above (from E. coli) or the mixture of GPa and GPb obtained from IMAC above (from Sf9 cells) is diluted 1:10 with the kinase assay buffer then mixed with the aforementioned phosphorylase kinase enzyme immobilized on the Affi-Gel beads. NaATP is added to 5 mM and MgCl2 to 6 mM. The resulting mixture is mixed gently at 25xc2x0 C. for 30 to 60 minutes. The sample is removed from the beads and the percent activation of GPb by conversion to GPa is estimated by determining GP enzyme activity in the presence and absence of 3.3 mM AMP. The percentage of total GP enzyme activity due to GPa enzyme activity (AMP-independent) is then calculated as follows:       %    ⁢          xe2x80x83        ⁢    of    ⁢          xe2x80x83        ⁢    total    ⁢          xe2x80x83        ⁢    HLGPa    =                    HLGP        ⁢                  xe2x80x83                ⁢        activity            -      AMP                      HLGP        ⁢                  xe2x80x83                ⁢        activity            +      AMP      
Alternately, the conversion of GPb to GPa can be monitored by isoelectric focusing, based on the shift in electrophoretic mobility that is noted following conversion of GPb to GPa. GP samples are analyzed by isoelectric focusing (IEF) utilizing the Pharmacia PfastGel System (Pharmacia Biotech. Inc., Piscataway, N.J.) using precast gels (pl range 4-6.5) and the manufacturer""s recommended method. The resolved GPa and GPb bands are then visualized on the gels by silver staining (2D-silver Stain II xe2x80x9cDaiichi Kitxe2x80x9d, Daiichi Pure Chemicals Co., LTD., Tokyo, Japan). Identification of GPa and GPb is made by comparison to E. coli derived GPa and GPb standards that are run in parallel on the same gels as the experimental samples.
B) GPa Activity Assay
The disease/condition treating/preventing activities described herein of the glycogen phosphorylase inhibitor compounds of this invention can be indirectly determined by assessing the effect of the compounds of this invention on the activity of the activated form of glycogen phosphorylase (GPa) by one of two methods; glycogen phosphorylase a activity is measured in the forward direction by monitoring the production of glucose-1-phosphate from glycogen or by following the reverse reaction, measuring glycogen synthesis from glucose-1-phosphate by the release of inorganic phosphate. All reactions can be run in triplicate in 96-well microtiter plates and the change in absorbance due to formation of the reaction product is measured at the wavelength specified below in a MCC/340 MKII Elisa Reader (Lab Systems, Finland), connected to a Titertech Microplate Stacker (ICN Biomedical Co, Huntsville, Ala.).
To measure the GPa enzyme activity in the forward direction, the production of glucose-1-phosphate from glycogen is monitored by the multienzyme coupled general method of Pesce et al. [Pesce, M. A., Bodourian, S. H., Harris, R. C. and Nicholson, J. F. (1977) Clinical Chemistry 23, 1711-1717] modified as follows: 1 to 100 xcexcg GPa, 10 units phosphoglucomutase and 15 units glucose-6-phosphate dehydrogenase (Boehringer Mannheim Biochemicals, Indianapolis, Ind.) are diluted to 1 mL in Buffer A (described hereinafter). Buffer A is at pH 7.2 and contains 50 mM HEPES, 100 mM KCl, 2.5 mM ethyleneglycoltetraacetic acid (EGTA), 2.5 mM MgCl2, 3.5 mM KH2PO4 and 0.5 mM dithiothreitol. 20 xcexcl of this stock is added to 80 xcexcl of Buffer A containing 0.47 mg/mL glycogen, 9.4 mM glucose, 0.63 mM of the oxidized form of nicotinamide adenine dinucleotide phosphate (NADP+). The compounds to be tested are added as 5 xcexcL of solution in 14% dimethylsulfoxide (DMSO). prior to the addition of the enzymes. The basal rate of GPa enzyme activity in the absence of inhibitors is determined by adding 5 xcexcL of 14% DMSO and a fully-inhibited rate of GPa enzyme activity is obtained by adding 20 xcexcL of 50 mM of the positive control test substance, caffeine. The reaction is followed at room temperature by measuring the conversion of oxidized NADP+ to reduced NADPH at 340 nm.
To measure the GPa enzyme activity in the reverse direction, the conversion of glucose-1-phosphate into glycogen plus inorganic phosphate is measured by the general method described by Engers et al. [Engers, H. D., Shechosky, S. and Madsen, N. B. (1970) Can. J. Biochem. 48, 746-754] modified as follows: 1 to 100 xcexcg GPa is diluted to 1 mL in Buffer B (described hereinafter). Buffer B is at pH 7.2 and contains 50 mM HEPES, 100 mM KCl, 2.5 mM EGTA, 2.5 mM MgCl2 and 0.5 mM dithiothreitol. 20 xcexcL of this stock is added to 80 xcexcL of Buffer B with 1.25 mg/mL glycogen, 9.4 mM glucose, and 0.63 mM glucose-1-phosphate. The compounds to be tested are added as 5 xcexcL of solution in 14% DMSO prior to the addition of the enzyme. The basal rate of GPa enzyme activity in the absence of added inhibitors is determined by adding 5 xcexcL of 14% DMSO and a fully-inhibited rate of GPa enzyme activity is obtained by adding 20 xcexcL of 50 mM caffeine. This mixture is incubated at room temperature for 1 hour and the inorganic phosphate released from the glucose-1-phosphate is measured by the general method of Lanzetta et al. [Lanzetta, P. A., Alvarez, L. J., Reinach, P. S. and Candia, O. A. (1979) Anal. Biochem. 100, 95-97] modified as follows: 150 xcexcL of 10 mg/mL ammonium molybdate, 0.38 mg/mL malachite green in 1 N HCl is added to 100 xcexcL of the enzyme mix. After a 20 minute incubation at room temperature, the absorbance is measured at 620 nm.
The above assays carried out with a range of concentrations of test compound allows the determination of an IC50 value (concentration of test compound required for 50% inhibition) for the in vitro inhibition of GPa enzyme activity by that test compound.
Administration of the compounds of this invention can be via any method which delivers a compound of this invention preferentially to the desired tissue (e.g., liver and/or cardiac tissues). These methods include oral routes, parenteral, intraduodenal routes, etc. Generally, the compounds of the present invention are administered in single (e.g., once daily) or multiple doses or via constant infusion.
The compounds of this invention are useful, for example, in reducing or minimizing damage effected directly to any tissue that may be susceptible to ischemia/reperfusion injury (e.g., heart, brain, lung, kidney, liver, gut, skeletal muscle, retina) as the result of an ischemic event (e.g., myocardial infarction). The active compound is therefore usefully employed prophylactically to prevent, i.e. (prospectively or prophylactically) to blunt or stem, tissue damage (e.g., myocardial tissue) in patients who are at risk for ischemia (e.g., myocardial ischemia).
Generally, the compounds of this invention are administered orally, or parenterally (e.g., intravenous, intramuscular, subcutaneous or intramedullary). Topical administration may also be indicated, for example, where the patient is suffering from gastrointestinal disorders or whenever the medication is best applied to the surface of a tissue or organ as determined by the attending physician.
The amount and timing of compounds administered will, of course, be dependent on the subject being treated, on the severity of the affliction, on the manner of administration and on the judgement of the prescribing physician. Thus, because of patient to patient variability, the dosages given below are a guideline and the physician may titrate doses of the drug to achieve the treatment that the physician considers appropriate for the patient. In considering the degree of treatment desired, the physician must balance a variety of factors such as age of the patient, presence of preexisting disease, as well as presence of other diseases (e.g., cardiovascular disease).
Thus, for example, in one mode of administration the compounds of this invention may be administered just prior to surgery (e.g., within twenty-four hours before surgery for example cardiac surgery) during or subsequent to surgery (e.g., within twenty-four hours after surgery) where there is risk of myocardial ischemia. The compounds of this invention may also be administered in a chronic daily mode.
An amount of the compounds of this invention is used that is effective for ischemic protection. A preferred dosage is about 0.001 to 100 mg/kg/day of the compound of this invention. An especially preferred dosage is about 0.01 to 50 mg/kg/day of the compound of this invention.
The compounds of the present invention are generally administered in the form of a pharmaceutical composition comprising at least one of the compounds of this invention together with a pharmaceutically acceptable vehicle or diluent. Thus, the compounds of this invention can be administered individually or together in any conventional oral, parenteral, rectal or transdermal dosage form.
For oral administration a pharmaceutical composition can take the form of solutions, suspensions, tablets, pills, capsules, powders, and the like. Tablets containing various excipients such as sodium citrate, calcium carbonate and calcium phosphate are employed along with various disintegrants such as starch and preferably potato or tapioca starch and certain complex silicates, together with binding agents such as polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often very useful for tabletting purposes. Solid compositions of a similar type are also employed as fillers in soft and hard-filled gelatin capsules; preferred materials in this connection also include lactose or milk sugar as well as high molecular weight polyethylene glycols. When aqueous suspensions and/or elixirs are desired for oral administration, the compounds of this invention can be combined with various sweetening agents, flavoring agents, coloring agents, emulsifying agents and/or suspending agents, as well as such diluents as water, ethanol, propylene glycol, glycerin and various like combinations thereof.
For purposes of parenteral administration, solutions, for example, in sesame or peanut oil or in aqueous propylene glycol can be employed, as well as sterile aqueous solutions of the corresponding water-soluble salts. Such aqueous solutions may be suitably buffered, if necessary, and the liquid diluent first rendered isotonic with sufficient saline or glucose. These aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal injection purposes. In this connection, the sterile aqueous media employed are all readily obtainable by standard techniques well-known to those skilled in the art.
For purposes of transdermal (e.g.,topical) administration, dilute sterile, aqueous or partially aqueous solutions (usually in about 0.1% to 5% concentration), otherwise similar to the above parenteral solutions, are prepared.
Methods of preparing various pharmaceutical compositions with a certain amount of active ingredient are known, or will be apparent in light of this disclosure, to those skilled in this art. For examples of methods of preparing pharmaceutical compositions, see Remington""s Pharmaceutical Sciences, Mack Publishing Company, Easter, Pa., 15th Edition (1975).
Pharmaceutical compositions according to the invention may contain for example 0.0001%-95% of the compound(s) of this invention. In any event, the composition or formulation to be administered will contain a quantity of a compound(s) according to the invention in an amount effective to treat the disease/condition of the subject being treated.
The two different compounds of this combination of this invention can be co-administered simultaneously or sequentially in any order, or as a single pharmaceutical composition comprising a compound of Formula I and an aldose reductase inhibitor as described above or a glycogen phosphorylase inhibitor as described above or a cardiovascular agent.
Since the present invention has an aspect that relates to the treatment of the disease/conditions described herein with a combination of active ingredients which may be administered separately, the invention also relates to combining separate pharmaceutical compositions in kit form. The kit comprises two separate pharmaceutical compositions: a compound of Formula I a prodrug thereof or a salt of such compound or prodrug and a second compound as described above. The kit comprises means for containing the separate compositions such as a container, a divided bottle or a divided foil packet. Typically the kit comprises directions for the administration of the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), are administered at different dosage intervals, or when titration of the individual components of the combination is desired by the prescribing physician.
An example of such a kit is a so-called blister pack. Blister packs are well known in the packaging industry and are being widely used for the packaging of pharmaceutical unit dosage forms (tablets, capsules, and the like). Blister packs generally consist of a sheet of relatively stiff material covered with a foil of a preferably transparent plastic material. During the packaging process recesses are formed in the plastic foil. The recesses have the size and shape of the tablets or capsules to be packed. Next, the tablets or capsules are placed in the recesses and the sheet of relatively stiff material is sealed against the plastic foil at the face of the foil which is opposite from the direction in which the recesses were formed. As a result, the tablets or capsules are sealed in the recesses between the plastic foil and the sheet. Preferably the strength of the sheet is such that the tablets or capsules can be removed from the blister pack by manually applying pressure on the recesses whereby an opening is formed in the sheet at the place of the recess. The tablet or capsule can then be removed via said opening.
It may be desirable to provide a memory aid on the kit, e.g., in the form of numbers next to the tablets or capsules whereby the numbers correspond with the days of the regimen which the tablets or capsules so specified should be ingested. Another example of such a memory aid is a calendar printed on the card, e.g., as follows xe2x80x9cFirst Week, Monday, Tuesday, . . . etc. . . . Second Week, Monday, Tuesday, . . . xe2x80x9d etc. Other variations of memory aids will be readily apparent. A xe2x80x9cdaily dosexe2x80x9d can be a single tablet or capsule or several pills or capsules to be taken on a given day. Also, a daily dose of Formula I compound can consist of one tablet or capsule while a daily dose of the second compound can consist of several tablets or capsules and vice versa. The memory aid should reflect this.
In another specific embodiment of the invention, a dispenser designed to dispense the daily doses one at a time in the order of their intended use is provided. Preferably, the dispenser is equipped with a memory-aid, so as to further facilitate compliance with the regimen. An example of such a memory-aid is a mechanical counter which indicates the number of daily doses that has been dispensed. Another example of such a memory-aid is a battery-powered micro-chip memory coupled with a liquid crystal readout, or audible reminder signal which, for example, reads out the date that the last daily dose has been taken and/or reminds one when the next dose is to be taken.
The compounds of this invention generally will be administered in a convenient formulation. The following formulation examples are illustrative only and are not intended to limit the scope of the present invention.
In the formulations which follow, xe2x80x9cactive ingredientxe2x80x9d means a compound(s) of this invention.
A tablet formulation is prepared using the ingredients below:
The components are blended and compressed to form tablets.
Alternatively, tablets each containing 0.25-100 mg of active ingredients are made up as follows:
The active ingredient, starch, and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a No. 14 mesh U.S. sieve. The granules so produced are dried at 50xc2x0-60xc2x0 C. and passed through a No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate, and talc, previously passed through a No. 60 U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets.
Suspensions each containing 0.25-100 mg of active ingredient per 5 ml dose are made as follows:
The active ingredient is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethyl cellulose and syrup to form smooth paste. The benzoic acid solution, flavor, and color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume. An aerosol solution is prepared containing the following ingredients:
The active ingredient is mixed with ethanol and the mixture added to a portion of the propellant 22, cooled to 30xc2x0 C., and transferred to a filling device. The required amount is then fed to a stainless steel container and diluted with the remaining propellant. The valve units are then fitted to the container.
Suppositories are prepared as follows:
The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimal necessary heat. The mixture is then poured into a suppository mold of nominal 2 g capacity and allowed to cool.
An intravenous formulation is prepared as follows:
The solution of the above ingredients is intravenously administered to a patient.
The active ingredient above may also be a combination of agents.
NMR spectra were recorded on a Varian XL-300 (Varian Co., Palo Alto, Calif.) a Bruker AM-300 spectrometer (Bruker Co., Billerica, Mass.) or a Varian Unity 400 at about 23xc2x0 C. at 300 or 400 MHz for proton. Chemical shifts are expressed in parts per million downfield from trimethylsilane. The peak shapes are denoted: as follows: s, singlet; d, doublet; t, triplet, q, quartet; m, multiplet; bs,=broad singlet. Resonances designated as exchangeable did not appear in a separate NMR experiment where the sample was shaken with several drops of D2O in the same solvent. Atmospheric pressure chemical ionization mass spectra (APCIMS) were obtained on a Fisons Platform II Spectrometer. Chemical ionization mass spectra (CIMS) were obtained on a Hewlett-Packard 5989 instrument (Hewlett-Packard Co., Palo Alto, Calif.) (ammonia ionization, PBMS). Where the intensity of chlorine or bromine-containing ions are described the expected intensity ratio was observed (approximately 3:1 for 35Cl/37Cl-containing ions and 1:1 for 79Br/81Br-containing ions) and M is based on 35Cl and 79Br. In some cases only representative 1H NMR and APCIMS peaks are given.
Column chromatography was performed with either Baker Silica Gel (40 xcexcm) (J. T. Baker, Phillipsburg, N.J.) or Silica Gel 60 (EM Sciences, Gibbstown, N.J.) in glass columns or in Flash 40(trademark) or Flash 12(trademark) (Biotage) (Charlottesville, Va.) columns under low nitrogen pressure. Radial Chromatography was performed using a Chromatron, (Harrison Research) (Palo Alto, Calif.) Unless otherwise specified, reagents were used as obtained from commercial sources. Dimethylformamide, 2-propanol, tetrahydrofuran, and dichloromethane used as reaction solvents were the anhydrous grade supplied by Aldrich Chemical Company (Milwaukee, Wis.). Microanalyses were performed by Schwarzkopf Microanalytical Laboratory, Woodside, N.Y. The terms xe2x80x9cconcentratedxe2x80x9d and xe2x80x9ccoevaporatedxe2x80x9d refer to removal of solvent at water aspirator pressure on a rotary evaporator with a bath temperature of less than 50xc2x0 C. Reactions conducted at xe2x80x9c0-20xc2x0 C.xe2x80x9d or xe2x80x9c0-25xc2x0 C.xe2x80x9d were conducted with initial cooling of the vessel in an insulated ice bath which was allowed to warm to room temperature over several hours. The abbreviation xe2x80x9cminxe2x80x9d and xe2x80x9chxe2x80x9d stand for xe2x80x9cminutesxe2x80x9d and xe2x80x9choursxe2x80x9d respectively.